CORD-19:8e3f518b27782ac9cc19e7556d74100ff23d972b JSONTXT 8 Projects

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Id Subject Object Predicate Lexical cue
TextSentencer_T1 0-117 Sentence denotes Porcine parvovirus infection activates mitochondria-mediated apoptotic signaling pathway by inducing ROS accumulation
TextSentencer_T1 0-117 Sentence denotes Porcine parvovirus infection activates mitochondria-mediated apoptotic signaling pathway by inducing ROS accumulation
TextSentencer_T2 119-127 Sentence denotes Abstract
TextSentencer_T2 119-127 Sentence denotes Abstract
TextSentencer_T3 128-139 Sentence denotes Background:
TextSentencer_T3 128-139 Sentence denotes Background:
TextSentencer_T4 140-262 Sentence denotes Porcine parvovirus (PPV) infection primarily causes reproductive failure of pregnant swine and results in host cell death.
TextSentencer_T4 140-262 Sentence denotes Porcine parvovirus (PPV) infection primarily causes reproductive failure of pregnant swine and results in host cell death.
TextSentencer_T5 263-327 Sentence denotes Boars, as an important disseminator, shed PPV to sows via semen.
TextSentencer_T5 263-327 Sentence denotes Boars, as an important disseminator, shed PPV to sows via semen.
TextSentencer_T6 328-434 Sentence denotes PPV infects and numerously replicates in boar testicle, which results in damage of swine testicle in vivo.
TextSentencer_T6 328-434 Sentence denotes PPV infects and numerously replicates in boar testicle, which results in damage of swine testicle in vivo.
TextSentencer_T7 435-554 Sentence denotes Reactive oxygen species (ROS), a mediator of cell apoptosis, play a crucial role in the mitochondria apoptotic pathway.
TextSentencer_T7 435-554 Sentence denotes Reactive oxygen species (ROS), a mediator of cell apoptosis, play a crucial role in the mitochondria apoptotic pathway.
TextSentencer_T8 555-651 Sentence denotes However, whether PPV infection induces ST cells apoptosis and ROS accumulation is still unclear.
TextSentencer_T8 555-651 Sentence denotes However, whether PPV infection induces ST cells apoptosis and ROS accumulation is still unclear.
TextSentencer_T9 652-660 Sentence denotes Methods:
TextSentencer_T9 652-660 Sentence denotes Methods:
TextSentencer_T10 661-840 Sentence denotes To determine the effects of PPV infection on the apoptosis, we detected morphological changes, DNA ladder, activities of caspases, and expression of PARP in PPV-infected ST cells.
TextSentencer_T10 661-840 Sentence denotes To determine the effects of PPV infection on the apoptosis, we detected morphological changes, DNA ladder, activities of caspases, and expression of PARP in PPV-infected ST cells.
TextSentencer_T11 841-1016 Sentence denotes Moreover, aiming to investigate the effect of PPV infection on the mitochondrial apoptotic pathway and ROS accumulation, we detected the Δψm, apoptosis-related genes, and ROS.
TextSentencer_T11 841-1016 Sentence denotes Moreover, aiming to investigate the effect of PPV infection on the mitochondrial apoptotic pathway and ROS accumulation, we detected the Δψm, apoptosis-related genes, and ROS.
TextSentencer_T12 1017-1163 Sentence denotes To investigate the role of ROS in the process of PPV-induced apoptosis, the ST cells were infected with PPV and treated with the ROS antioxidants.
TextSentencer_T12 1017-1163 Sentence denotes To investigate the role of ROS in the process of PPV-induced apoptosis, the ST cells were infected with PPV and treated with the ROS antioxidants.
TextSentencer_T13 1164-1353 Sentence denotes The ROS level was measured using Reactive Oxygen Species Assay Kit and the Δψm, expression level of Bcl-2, translocation of Bax, and redistribution of mitochondria cytochrome c were tested.
TextSentencer_T13 1164-1353 Sentence denotes The ROS level was measured using Reactive Oxygen Species Assay Kit and the Δψm, expression level of Bcl-2, translocation of Bax, and redistribution of mitochondria cytochrome c were tested.
TextSentencer_T14 1354-1521 Sentence denotes In this study, we demonstrated that PPV infection could induce apoptosis that was characterized by morphological changes, DNA fragmentation and activation of caspases.
TextSentencer_T14 1354-1521 Sentence denotes In this study, we demonstrated that PPV infection could induce apoptosis that was characterized by morphological changes, DNA fragmentation and activation of caspases.
TextSentencer_T15 1522-1824 Sentence denotes Moreover, PPV infection suppressed Bcl-2 expression, enhanced Bax expression and translocation to mitochondria, decreased the mitochondrial transmembrane potential, and triggered the release of cytochrome c, which caused the subsequent activation of caspase-9 and caspase-3 and initiation of apoptosis.
TextSentencer_T15 1522-1824 Sentence denotes Moreover, PPV infection suppressed Bcl-2 expression, enhanced Bax expression and translocation to mitochondria, decreased the mitochondrial transmembrane potential, and triggered the release of cytochrome c, which caused the subsequent activation of caspase-9 and caspase-3 and initiation of apoptosis.
TextSentencer_T16 1825-1932 Sentence denotes However, during the process of PPV-induced apoptosis, the protein levels of Fas and FasL were not affected.
TextSentencer_T16 1825-1932 Sentence denotes However, during the process of PPV-induced apoptosis, the protein levels of Fas and FasL were not affected.
TextSentencer_T17 1933-1999 Sentence denotes Further studies showed that PPV infection caused ROS accumulation.
TextSentencer_T17 1933-1999 Sentence denotes Further studies showed that PPV infection caused ROS accumulation.
TextSentencer_T18 2000-2191 Sentence denotes Inhibition of ROS could reduce mitochondrial transmembrane potential and could significantly block ST cells apoptosis via suppressing Bax translocation, cytochrome c and caspase-3 activation.
TextSentencer_T18 2000-2191 Sentence denotes Inhibition of ROS could reduce mitochondrial transmembrane potential and could significantly block ST cells apoptosis via suppressing Bax translocation, cytochrome c and caspase-3 activation.
TextSentencer_T19 2192-2204 Sentence denotes Conclusions:
TextSentencer_T19 2192-2204 Sentence denotes Conclusions:
TextSentencer_T20 2205-2378 Sentence denotes All these results suggest that PPV-induced ROS accumulation mediates apoptosis in ST cells, which provided theoretical basis for the molecular pathogenesis of PPV infection.
TextSentencer_T20 2205-2378 Sentence denotes All these results suggest that PPV-induced ROS accumulation mediates apoptosis in ST cells, which provided theoretical basis for the molecular pathogenesis of PPV infection.
TextSentencer_T21 2380-2470 Sentence denotes PPV is a member of parvoviridae, containing a negative single-stranded DNA genome [1, 2] .
TextSentencer_T21 2380-2470 Sentence denotes PPV is a member of parvoviridae, containing a negative single-stranded DNA genome [1, 2] .
TextSentencer_T22 2471-2616 Sentence denotes PPV infection may lead to reproductive failure, including infertility, embryonic death, stillbirth and fetal mummification in pregnant sows [3] .
TextSentencer_T22 2471-2616 Sentence denotes PPV infection may lead to reproductive failure, including infertility, embryonic death, stillbirth and fetal mummification in pregnant sows [3] .
TextSentencer_T23 2617-2773 Sentence denotes PPV infection mainly causes reproductive clinical syndrome, including infertility, abortion, stillbirth, neonatal death, and reduced neonatal vitality [4] .
TextSentencer_T23 2617-2773 Sentence denotes PPV infection mainly causes reproductive clinical syndrome, including infertility, abortion, stillbirth, neonatal death, and reduced neonatal vitality [4] .
TextSentencer_T24 2774-2827 Sentence denotes Boars play an important role in dissemination of PPV.
TextSentencer_T24 2774-2827 Sentence denotes Boars play an important role in dissemination of PPV.
TextSentencer_T25 2828-2917 Sentence denotes It was reported that PPV infection damages both uterine [5] and testicles [6, 7] in vivo.
TextSentencer_T25 2828-2917 Sentence denotes It was reported that PPV infection damages both uterine [5] and testicles [6, 7] in vivo.
TextSentencer_T26 2918-3052 Sentence denotes Swine testicular is the reproductive organ and responsible for generating semen, which serve as one of the transmission routes of PPV.
TextSentencer_T26 2918-3052 Sentence denotes Swine testicular is the reproductive organ and responsible for generating semen, which serve as one of the transmission routes of PPV.
TextSentencer_T27 3053-3140 Sentence denotes Therefore, we use ST cells to investigate the mechanism of PPV infection in this study.
TextSentencer_T27 3053-3140 Sentence denotes Therefore, we use ST cells to investigate the mechanism of PPV infection in this study.
TextSentencer_T28 3141-3332 Sentence denotes The infection of animal virus such as human parvovirus [8, 9] , canine parvovirus [10] may induce host cells apoptosis and is a major factor of cell death and tissue damage in viral diseases.
TextSentencer_T28 3141-3332 Sentence denotes The infection of animal virus such as human parvovirus [8, 9] , canine parvovirus [10] may induce host cells apoptosis and is a major factor of cell death and tissue damage in viral diseases.
TextSentencer_T29 3333-3433 Sentence denotes Previous studies indicated that PPV infection caused apoptosis via p53 pathway in PK-15 cells [11] .
TextSentencer_T29 3333-3433 Sentence denotes Previous studies indicated that PPV infection caused apoptosis via p53 pathway in PK-15 cells [11] .
TextSentencer_T30 3434-3595 Sentence denotes Although, we previously demonstrated that PPV induced apoptosis PK-15 cells, whether PPV induces ST cells apoptosis and the role of ROS in apoptosis are unknown.
TextSentencer_T30 3434-3595 Sentence denotes Although, we previously demonstrated that PPV induced apoptosis PK-15 cells, whether PPV induces ST cells apoptosis and the role of ROS in apoptosis are unknown.
TextSentencer_T31 3596-3685 Sentence denotes Apoptosis, a programmed cell death (PCD), is an important pathological cellular response.
TextSentencer_T31 3596-3685 Sentence denotes Apoptosis, a programmed cell death (PCD), is an important pathological cellular response.
TextSentencer_T32 3686-3792 Sentence denotes In vertebrate cells, the apoptotic pathways mainly include the extrinsic and the intrinsic pathways [12] .
TextSentencer_T32 3686-3792 Sentence denotes In vertebrate cells, the apoptotic pathways mainly include the extrinsic and the intrinsic pathways [12] .
TextSentencer_T33 3793-3948 Sentence denotes The extrinsic apoptotic pathway is induced by stimulation of death receptors that belongs to the tumor necrosis factor receptor (TNFR) family, such as Fas.
TextSentencer_T33 3793-3948 Sentence denotes The extrinsic apoptotic pathway is induced by stimulation of death receptors that belongs to the tumor necrosis factor receptor (TNFR) family, such as Fas.
TextSentencer_T34 3949-4080 Sentence denotes The induction of intrinsic apoptotic pathway is associated with mitochondria upon activation that is highly regulated by ROS [13] .
TextSentencer_T34 3949-4080 Sentence denotes The induction of intrinsic apoptotic pathway is associated with mitochondria upon activation that is highly regulated by ROS [13] .
TextSentencer_T35 4081-4230 Sentence denotes ROS, known as toxic products of cellular metabolism, are mainly produced by the mitochondria in most vertebrate cells and act as signaling molecules.
TextSentencer_T35 4081-4230 Sentence denotes ROS, known as toxic products of cellular metabolism, are mainly produced by the mitochondria in most vertebrate cells and act as signaling molecules.
TextSentencer_T36 4231-4366 Sentence denotes Moreover, growing evidence suggests ROS are involved in regulation of many pathological processes such as cellular apoptosis [14, 15] .
TextSentencer_T36 4231-4366 Sentence denotes Moreover, growing evidence suggests ROS are involved in regulation of many pathological processes such as cellular apoptosis [14, 15] .
TextSentencer_T37 4367-4447 Sentence denotes However excessive ROS is toxic to cells, especially in the process of apoptosis.
TextSentencer_T37 4367-4447 Sentence denotes However excessive ROS is toxic to cells, especially in the process of apoptosis.
TextSentencer_T38 4448-4667 Sentence denotes It is indicated that ROS accumulation was able to enhance apoptosis by collapsing the mitochondrial potential, inducing mitochondrial oxidation channel, and releasing cytochrome C from mitochondria to the cytosol [16] .
TextSentencer_T38 4448-4667 Sentence denotes It is indicated that ROS accumulation was able to enhance apoptosis by collapsing the mitochondrial potential, inducing mitochondrial oxidation channel, and releasing cytochrome C from mitochondria to the cytosol [16] .
TextSentencer_T39 4668-4727 Sentence denotes ROS accumulation is also linked to virus-induced apoptosis.
TextSentencer_T39 4668-4727 Sentence denotes ROS accumulation is also linked to virus-induced apoptosis.
TextSentencer_T40 4728-4826 Sentence denotes For instance, parvovirus H-1 infection induces apoptosis via mediating ROS accumulation [17, 18] .
TextSentencer_T40 4728-4826 Sentence denotes For instance, parvovirus H-1 infection induces apoptosis via mediating ROS accumulation [17, 18] .
TextSentencer_T41 4827-4947 Sentence denotes We have reported that PPV infection could be able to trigger cell apoptosis through mitochondria-mediated pathway [19] .
TextSentencer_T41 4827-4947 Sentence denotes We have reported that PPV infection could be able to trigger cell apoptosis through mitochondria-mediated pathway [19] .
TextSentencer_T42 4948-5028 Sentence denotes However, whether ROS are associated with PPV-induced apoptosis is still unclear.
TextSentencer_T42 4948-5028 Sentence denotes However, whether ROS are associated with PPV-induced apoptosis is still unclear.
TextSentencer_T43 5029-5131 Sentence denotes In this study, we asked whether PPV infection could induce apoptosis via ROS accumulation in ST cells.
TextSentencer_T43 5029-5131 Sentence denotes In this study, we asked whether PPV infection could induce apoptosis via ROS accumulation in ST cells.
TextSentencer_T44 5132-5218 Sentence denotes To address this issue, we detected whether PPV infection causes apoptosis in ST cells.
TextSentencer_T44 5132-5218 Sentence denotes To address this issue, we detected whether PPV infection causes apoptosis in ST cells.
TextSentencer_T45 5219-5307 Sentence denotes In addition, the ROS induced by PPV infection was assessed compared with mock infection.
TextSentencer_T45 5219-5307 Sentence denotes In addition, the ROS induced by PPV infection was assessed compared with mock infection.
TextSentencer_T46 5308-5412 Sentence denotes The results showed that PPV infection could triggers ST cell apoptosis via PPV-induced ROS accumulation.
TextSentencer_T46 5308-5412 Sentence denotes The results showed that PPV infection could triggers ST cell apoptosis via PPV-induced ROS accumulation.
TextSentencer_T47 5413-5539 Sentence denotes To determine the role of PPV infection in apoptosis of ST cells, we observed the morphological change of PPVinfected ST cells.
TextSentencer_T47 5413-5539 Sentence denotes To determine the role of PPV infection in apoptosis of ST cells, we observed the morphological change of PPVinfected ST cells.
TextSentencer_T48 5540-5798 Sentence denotes PPV-infected cells started to show typical apoptotic features including chromatin condensation (Fig. 1a , indicated by yellow arrows) after 24 hpi and nuclear fragmentation compared with mock-infected cells (Fig. 1a , indicated by white arrows) after 36 hpi.
TextSentencer_T48 5540-5798 Sentence denotes PPV-infected cells started to show typical apoptotic features including chromatin condensation (Fig. 1a , indicated by yellow arrows) after 24 hpi and nuclear fragmentation compared with mock-infected cells (Fig. 1a , indicated by white arrows) after 36 hpi.
TextSentencer_T49 5799-5952 Sentence denotes Next, we detected the pattern of chromosomal DNA fragmentation and found that DNA ladders appeared at 48 hpi in PPV infected cells at 1.0 MOI (Fig. 1b) .
TextSentencer_T49 5799-5952 Sentence denotes Next, we detected the pattern of chromosomal DNA fragmentation and found that DNA ladders appeared at 48 hpi in PPV infected cells at 1.0 MOI (Fig. 1b) .
TextSentencer_T50 5953-6011 Sentence denotes Then the apoptotic rate was assessed using flow cytometry.
TextSentencer_T50 5953-6011 Sentence denotes Then the apoptotic rate was assessed using flow cytometry.
TextSentencer_T51 6012-6169 Sentence denotes As shown in Fig. 1c , the apoptotic rate of PPV-infected cells was increased at 24 hpi in comparison with the mockinfected cells and appeared time-dependent.
TextSentencer_T51 6012-6169 Sentence denotes As shown in Fig. 1c , the apoptotic rate of PPV-infected cells was increased at 24 hpi in comparison with the mockinfected cells and appeared time-dependent.
TextSentencer_T52 6170-6331 Sentence denotes To investigate the mechanism of apoptosis induced by PPV infection, we further examined the activities of several vital caspases, including caspase-3, -8 and -9.
TextSentencer_T52 6170-6331 Sentence denotes To investigate the mechanism of apoptosis induced by PPV infection, we further examined the activities of several vital caspases, including caspase-3, -8 and -9.
TextSentencer_T53 6332-6456 Sentence denotes The results indicated that the caspase-9 and -3 were activated after PPV infection at 1.0 MOI as early as 12 hpi (Fig. 1d ).
TextSentencer_T53 6332-6456 Sentence denotes The results indicated that the caspase-9 and -3 were activated after PPV infection at 1.0 MOI as early as 12 hpi (Fig. 1d ).
TextSentencer_T54 6457-6525 Sentence denotes However the activity of caspase-8 was not affected by PPV infection.
TextSentencer_T54 6457-6525 Sentence denotes However the activity of caspase-8 was not affected by PPV infection.
TextSentencer_T55 6526-6696 Sentence denotes In addition, poly-ADP-ribose polymerase (PARP), an indicator of caspase-3 activation, was cleaved in PPV-infected ST cells at 12 hpi in a timedependent manner (Fig. 1e ).
TextSentencer_T55 6526-6696 Sentence denotes In addition, poly-ADP-ribose polymerase (PARP), an indicator of caspase-3 activation, was cleaved in PPV-infected ST cells at 12 hpi in a timedependent manner (Fig. 1e ).
TextSentencer_T56 6697-6757 Sentence denotes In addition, the Fas and FasL were not affected (Fig. 1f ) .
TextSentencer_T56 6697-6757 Sentence denotes In addition, the Fas and FasL were not affected (Fig. 1f ) .
TextSentencer_T57 6758-6890 Sentence denotes Taken together, these results indicated that PPV infection induced apoptosis in ST cells through activating caspase-9 and caspase-3.
TextSentencer_T57 6758-6890 Sentence denotes Taken together, these results indicated that PPV infection induced apoptosis in ST cells through activating caspase-9 and caspase-3.
TextSentencer_T58 6891-6952 Sentence denotes Δψm plays an important role in the process of cell apoptosis.
TextSentencer_T58 6891-6952 Sentence denotes Δψm plays an important role in the process of cell apoptosis.
TextSentencer_T59 6953-7048 Sentence denotes Once the Δψm is collapsed, mitochondria-mediated apoptotic signaling pathway will be activated.
TextSentencer_T59 6953-7048 Sentence denotes Once the Δψm is collapsed, mitochondria-mediated apoptotic signaling pathway will be activated.
TextSentencer_T60 7049-7161 Sentence denotes To determine the effect of PPV infection on Δψm, we detected the Δψm of PPV-infected and mock-infected ST cells.
TextSentencer_T60 7049-7161 Sentence denotes To determine the effect of PPV infection on Δψm, we detected the Δψm of PPV-infected and mock-infected ST cells.
TextSentencer_T61 7162-7298 Sentence denotes As shown in Fig. 2a , the percentage of Δψm-depolarized cells increased 6 % at 6 hpi and the level of depolarization was time-dependent.
TextSentencer_T61 7162-7298 Sentence denotes As shown in Fig. 2a , the percentage of Δψm-depolarized cells increased 6 % at 6 hpi and the level of depolarization was time-dependent.
TextSentencer_T62 7299-7449 Sentence denotes During mitochondria-mediated apoptosis, Δψm was regulated by Bcl-2 family members, such as pro-apoptotic protein Bax and anti-apoptotic protein Bcl-2.
TextSentencer_T62 7299-7449 Sentence denotes During mitochondria-mediated apoptosis, Δψm was regulated by Bcl-2 family members, such as pro-apoptotic protein Bax and anti-apoptotic protein Bcl-2.
TextSentencer_T63 7450-7544 Sentence denotes To explore the effects of PPV infection on Bcl-2 and Bax, western blot analysis was performed.
TextSentencer_T63 7450-7544 Sentence denotes To explore the effects of PPV infection on Bcl-2 and Bax, western blot analysis was performed.
TextSentencer_T64 7545-7664 Sentence denotes The western blot results showed that PPV infection increased Bax expression and suppressed Bcl-2 expression (Fig. 2b) .
TextSentencer_T64 7545-7664 Sentence denotes The western blot results showed that PPV infection increased Bax expression and suppressed Bcl-2 expression (Fig. 2b) .
TextSentencer_T65 7665-7910 Sentence denotes To further detect whether the collapse of Δψm is associated with the release of cytochrome c and the translocation of Bax, the cytoplasmic and mitochondrial proteins were analyzed by western blot and respectively normalized to β-actin and Cox-4.
TextSentencer_T65 7665-7910 Sentence denotes To further detect whether the collapse of Δψm is associated with the release of cytochrome c and the translocation of Bax, the cytoplasmic and mitochondrial proteins were analyzed by western blot and respectively normalized to β-actin and Cox-4.
TextSentencer_T66 7911-8131 Sentence denotes The results showed that PPV-infection promoted the translocation of Bax from the cytosol to the mitochondria and enhanced the release of mitochondrial cytochrome c from mitochondria to cytosol at 12 hpi ( Fig. 2c and d).
TextSentencer_T66 7911-8131 Sentence denotes The results showed that PPV-infection promoted the translocation of Bax from the cytosol to the mitochondria and enhanced the release of mitochondrial cytochrome c from mitochondria to cytosol at 12 hpi ( Fig. 2c and d).
TextSentencer_T67 8132-8237 Sentence denotes All these results indicated that PPV infection activated the mitochondrial apoptotic pathway in ST cells.
TextSentencer_T67 8132-8237 Sentence denotes All these results indicated that PPV infection activated the mitochondrial apoptotic pathway in ST cells.
TextSentencer_T68 8238-8365 Sentence denotes ROS play an important role in the process of apoptosis [16, 17] and activate the mitochondria-mediated apoptotic pathway [19] .
TextSentencer_T68 8238-8365 Sentence denotes ROS play an important role in the process of apoptosis [16, 17] and activate the mitochondria-mediated apoptotic pathway [19] .
TextSentencer_T69 8366-8550 Sentence denotes To investigate the role of ROS in the process of PPV-induced apoptosis, we detected ROS level in PPV-infected cells using DCFH-DA followed by observation under fluorescence microscopy.
TextSentencer_T69 8366-8550 Sentence denotes To investigate the role of ROS in the process of PPV-induced apoptosis, we detected ROS level in PPV-infected cells using DCFH-DA followed by observation under fluorescence microscopy.
TextSentencer_T70 8551-8702 Sentence denotes The results indicated that the fluorescence reached to the highest level at 24 hpi, and then decreased gradually along with the time course (Fig. 3a) .
TextSentencer_T70 8551-8702 Sentence denotes The results indicated that the fluorescence reached to the highest level at 24 hpi, and then decreased gradually along with the time course (Fig. 3a) .
TextSentencer_T71 8703-8827 Sentence denotes To further demonstrate the ROS level, quantitative analysis of ROS in PPV-infected cells was performed using flow cytometry.
TextSentencer_T71 8703-8827 Sentence denotes To further demonstrate the ROS level, quantitative analysis of ROS in PPV-infected cells was performed using flow cytometry.
TextSentencer_T72 8828-9015 Sentence denotes As shown in Fig. 3b and c, ROS was significantly accumulated at 24 hpi in comparison with mock-infected cells and subsequently decreased at 36 hpi continuously with the extension of time.
TextSentencer_T72 8828-9015 Sentence denotes As shown in Fig. 3b and c, ROS was significantly accumulated at 24 hpi in comparison with mock-infected cells and subsequently decreased at 36 hpi continuously with the extension of time.
TextSentencer_T73 9016-9101 Sentence denotes All the data indicated that PPV infection could trigger ROS accumulation in ST cells.
TextSentencer_T73 9016-9101 Sentence denotes All the data indicated that PPV infection could trigger ROS accumulation in ST cells.
TextSentencer_T74 9102-9198 Sentence denotes ROS accumulation may initiate cell apoptosis via activating the intrinsic mitochondrial pathway.
TextSentencer_T74 9102-9198 Sentence denotes ROS accumulation may initiate cell apoptosis via activating the intrinsic mitochondrial pathway.
TextSentencer_T75 9199-9332 Sentence denotes Moreover, excessive ROS also leads to the damage of mitochondrial DNA and the permeabilization of mitochondrial outer membrane [20] .
TextSentencer_T75 9199-9332 Sentence denotes Moreover, excessive ROS also leads to the damage of mitochondrial DNA and the permeabilization of mitochondrial outer membrane [20] .
TextSentencer_T76 9333-9582 Sentence denotes To further explore the effect of ROS on Δψm,we used ROS antioxidants PDTC (100 μM) (Merck, Germany) and NAC (100 μM) (Sigma-Aldrich, US) to suppress ROS production in ST cells infected with PPV at 1 MOI and 24 hpi and measured the ROS level and Δψm.
TextSentencer_T76 9333-9582 Sentence denotes To further explore the effect of ROS on Δψm,we used ROS antioxidants PDTC (100 μM) (Merck, Germany) and NAC (100 μM) (Sigma-Aldrich, US) to suppress ROS production in ST cells infected with PPV at 1 MOI and 24 hpi and measured the ROS level and Δψm.
TextSentencer_T77 9583-9780 Sentence denotes Both PDTC and NAC, which did not show cytotoxicity at concentrations used (data not shown), significantly prevented the production of ROS (Fig. 4a) and reduced Δψm in PPV-infected cells (Fig. 4b) .
TextSentencer_T77 9583-9780 Sentence denotes Both PDTC and NAC, which did not show cytotoxicity at concentrations used (data not shown), significantly prevented the production of ROS (Fig. 4a) and reduced Δψm in PPV-infected cells (Fig. 4b) .
TextSentencer_T78 9781-9902 Sentence denotes These results indicated that PPV infection decreased mitochondrial membrane potential through inducting ROS accumulation.
TextSentencer_T78 9781-9902 Sentence denotes These results indicated that PPV infection decreased mitochondrial membrane potential through inducting ROS accumulation.
TextSentencer_T79 9903-10157 Sentence denotes To further investigate the role of ROS accumulation in apoptosis, we analyzed the expression of Bcl-2, translocation of Bax, and the redistribution of mitochondria cytochrome c in ST cells infected with PPV at 1 MOI, 24 hpi and treated with PDTC and NAC.
TextSentencer_T79 9903-10157 Sentence denotes To further investigate the role of ROS accumulation in apoptosis, we analyzed the expression of Bcl-2, translocation of Bax, and the redistribution of mitochondria cytochrome c in ST cells infected with PPV at 1 MOI, 24 hpi and treated with PDTC and NAC.
TextSentencer_T80 10158-10436 Sentence denotes As shown in Fig. 4c , the decrease of Bcl-2, the translocation of Bax from cytoplasm to mitochondria, and the release of mitochondrial cytochrome c to cytoplasm in PPV infected ST cells were blocked by PDTC and NAC treatment in comparison with these in PPV-infected cells alone.
TextSentencer_T80 10158-10436 Sentence denotes As shown in Fig. 4c , the decrease of Bcl-2, the translocation of Bax from cytoplasm to mitochondria, and the release of mitochondrial cytochrome c to cytoplasm in PPV infected ST cells were blocked by PDTC and NAC treatment in comparison with these in PPV-infected cells alone.
TextSentencer_T81 10437-10616 Sentence denotes In addition, caspase-3 activity and apoptotic rate were remarkably decreased in the ST cells infected with PPV at 1 MOI and 24 hpi in the presence of PDTC or NAC (Fig. 4d and e) .
TextSentencer_T81 10437-10616 Sentence denotes In addition, caspase-3 activity and apoptotic rate were remarkably decreased in the ST cells infected with PPV at 1 MOI and 24 hpi in the presence of PDTC or NAC (Fig. 4d and e) .
TextSentencer_T82 10617-10739 Sentence denotes The data indicated that the accumulation of ROS induced by PPV infection contributed to the apoptosis at 1 MOI and 24 hpi.
TextSentencer_T82 10617-10739 Sentence denotes The data indicated that the accumulation of ROS induced by PPV infection contributed to the apoptosis at 1 MOI and 24 hpi.
TextSentencer_T83 10740-10961 Sentence denotes Apoptosis plays a predominant role in contributing to the pathogenic process of virus infection [21] and presents specific morphological and molecular changes, including chromatin margination, DNA fragmentation [22, 23] .
TextSentencer_T83 10740-10961 Sentence denotes Apoptosis plays a predominant role in contributing to the pathogenic process of virus infection [21] and presents specific morphological and molecular changes, including chromatin margination, DNA fragmentation [22, 23] .
TextSentencer_T84 10962-11069 Sentence denotes Previous studies showed that PPV infection induced PK-15 apoptosis via mitochondria-mediated pathway [19] .
TextSentencer_T84 10962-11069 Sentence denotes Previous studies showed that PPV infection induced PK-15 apoptosis via mitochondria-mediated pathway [19] .
TextSentencer_T85 11070-11233 Sentence denotes Evidence shows that oxidative stress is an important regulatory factor of virus-induced apoptosis and induced by various triggers, including viral infection [24] .
TextSentencer_T85 11070-11233 Sentence denotes Evidence shows that oxidative stress is an important regulatory factor of virus-induced apoptosis and induced by various triggers, including viral infection [24] .
TextSentencer_T86 11234-11437 Sentence denotes In this study, we found that PPV infection induced apoptosis of ST cells via activating mitochondrialmediated pathway and PPV-induced ROS accumulation was involved in regulating the process of apoptosis.
TextSentencer_T86 11234-11437 Sentence denotes In this study, we found that PPV infection induced apoptosis of ST cells via activating mitochondrialmediated pathway and PPV-induced ROS accumulation was involved in regulating the process of apoptosis.
TextSentencer_T87 11438-11560 Sentence denotes Apoptosis is primarily classified into two pathways regulating cell death involving the continuous activation of caspases.
TextSentencer_T87 11438-11560 Sentence denotes Apoptosis is primarily classified into two pathways regulating cell death involving the continuous activation of caspases.
TextSentencer_T88 11561-11804 Sentence denotes Of these, the extrinsic pathway participates in the ligation of particular "death" receptors (e.g., Fas) by recruitment of adaptor molecules to form the deathinducing signaling complex (DISC) and activate upstream caspases via caspase-8 [25] .
TextSentencer_T88 11561-11804 Sentence denotes Of these, the extrinsic pathway participates in the ligation of particular "death" receptors (e.g., Fas) by recruitment of adaptor molecules to form the deathinducing signaling complex (DISC) and activate upstream caspases via caspase-8 [25] .
TextSentencer_T89 11805-11978 Sentence denotes The mitochondrion-mediated (intrinsic) pathway is characterized by changed mitochondrial membrane potential and release of cytochrome c from mitochondria into the cytoplasm.
TextSentencer_T89 11805-11978 Sentence denotes The mitochondrion-mediated (intrinsic) pathway is characterized by changed mitochondrial membrane potential and release of cytochrome c from mitochondria into the cytoplasm.
TextSentencer_T90 11979-12154 Sentence denotes In the cytoplasm, cytochrome c then is involved in the formation of a multiprotein complex known as the "apoptosome" and activates the caspase cascade through caspase-9 [26] .
TextSentencer_T90 11979-12154 Sentence denotes In the cytoplasm, cytochrome c then is involved in the formation of a multiprotein complex known as the "apoptosome" and activates the caspase cascade through caspase-9 [26] .
TextSentencer_T91 12155-12273 Sentence denotes In this study, we determined caspase-3 and caspase-9 were activated by PPV infection, but caspase-8 was not activated.
TextSentencer_T91 12155-12273 Sentence denotes In this study, we determined caspase-3 and caspase-9 were activated by PPV infection, but caspase-8 was not activated.
TextSentencer_T92 12274-12376 Sentence denotes This suggests that PPV infection induced the apoptosis of ST cells via mitochondrion-mediated pathway.
TextSentencer_T92 12274-12376 Sentence denotes This suggests that PPV infection induced the apoptosis of ST cells via mitochondrion-mediated pathway.
TextSentencer_T93 12377-12609 Sentence denotes Previous studies indicated that Bcl-2 family proteins as major regulators, including Bcl-2 and Bax, regulated the integrity of mitochondrial membranes, and the destruction of the mitochondrial membrane potential leaded to [27, 28] .
TextSentencer_T93 12377-12609 Sentence denotes Previous studies indicated that Bcl-2 family proteins as major regulators, including Bcl-2 and Bax, regulated the integrity of mitochondrial membranes, and the destruction of the mitochondrial membrane potential leaded to [27, 28] .
TextSentencer_T94 12610-12788 Sentence denotes Apoptosis can be triggered by the decreased mitochondrial permeability, which causes the release of apoptotic initiating factors, and the activation of caspase such as caspase-3.
TextSentencer_T94 12610-12788 Sentence denotes Apoptosis can be triggered by the decreased mitochondrial permeability, which causes the release of apoptotic initiating factors, and the activation of caspase such as caspase-3.
TextSentencer_T95 12789-12905 Sentence denotes Some parvoviruses infection may lead to the increase of membrane permeability, subsequently inducing cell apoptosis.
TextSentencer_T95 12789-12905 Sentence denotes Some parvoviruses infection may lead to the increase of membrane permeability, subsequently inducing cell apoptosis.
TextSentencer_T96 12906-13055 Sentence denotes For example, bocavirus minute virus of canines (MVC) results in the time-dependent disruption of mitochondrial outer membrane potential (MOMP) [29] .
TextSentencer_T96 12906-13055 Sentence denotes For example, bocavirus minute virus of canines (MVC) results in the time-dependent disruption of mitochondrial outer membrane potential (MOMP) [29] .
TextSentencer_T97 13056-13251 Sentence denotes Here we showed that mitochondrial membrane potential was significantly reduced in manner of time-dependent, which is consistent with the finding that MVC infection induced the disruption of MOMP.
TextSentencer_T97 13056-13251 Sentence denotes Here we showed that mitochondrial membrane potential was significantly reduced in manner of time-dependent, which is consistent with the finding that MVC infection induced the disruption of MOMP.
TextSentencer_T98 13252-13425 Sentence denotes The oxidative damage also leads to the release of cellular toxic proteins such as cytochrome c from the mitochondria to the cytosol, subsequently triggering apoptosis [14] .
TextSentencer_T98 13252-13425 Sentence denotes The oxidative damage also leads to the release of cellular toxic proteins such as cytochrome c from the mitochondria to the cytosol, subsequently triggering apoptosis [14] .
TextSentencer_T99 13426-13581 Sentence denotes To explore whether ROS was involved in the apoptosis induced by PPV infection, we investigated the direct effect of PPV infection on the oxidative profile.
TextSentencer_T99 13426-13581 Sentence denotes To explore whether ROS was involved in the apoptosis induced by PPV infection, we investigated the direct effect of PPV infection on the oxidative profile.
TextSentencer_T100 13582-13737 Sentence denotes Time course analysis of PPV infected ST cells showed that ROS accumulation increased significantly, suggesting that PPV infection induced ROS accumulation.
TextSentencer_T100 13582-13737 Sentence denotes Time course analysis of PPV infected ST cells showed that ROS accumulation increased significantly, suggesting that PPV infection induced ROS accumulation.
TextSentencer_T101 13738-13911 Sentence denotes In this study, two ROS inhibitors, PDTC and NAC, significantly attenuated PPV-induced Bax translocation, the release of cytochrome c, caspase-3 activity, and apoptotic rate.
TextSentencer_T101 13738-13911 Sentence denotes In this study, two ROS inhibitors, PDTC and NAC, significantly attenuated PPV-induced Bax translocation, the release of cytochrome c, caspase-3 activity, and apoptotic rate.
TextSentencer_T102 13912-14016 Sentence denotes The data indicate that ROS accumulation induced by PPV infection increases mitochondrial outer membrane.
TextSentencer_T102 13912-14016 Sentence denotes The data indicate that ROS accumulation induced by PPV infection increases mitochondrial outer membrane.
TextSentencer_T103 14017-14231 Sentence denotes Our finding is consistent with that the canine parvovirus (CPV) and rat parvovirus H-1 (H-1PV) nonstructureal NS1 protein induced apoptosis via accumulation of intracellular reactive oxygen species (ROS) [18, 30] .
TextSentencer_T103 14017-14231 Sentence denotes Our finding is consistent with that the canine parvovirus (CPV) and rat parvovirus H-1 (H-1PV) nonstructureal NS1 protein induced apoptosis via accumulation of intracellular reactive oxygen species (ROS) [18, 30] .
TextSentencer_T104 14232-14307 Sentence denotes Our data provide a link between ROS and apoptosis induced by PPV infection.
TextSentencer_T104 14232-14307 Sentence denotes Our data provide a link between ROS and apoptosis induced by PPV infection.
TextSentencer_T105 14308-14457 Sentence denotes The results demonstrated that PPV infection induced ST cells apoptosis via activating ROS accumulation and mitochondria-mediated apoptotic signaling.
TextSentencer_T105 14308-14457 Sentence denotes The results demonstrated that PPV infection induced ST cells apoptosis via activating ROS accumulation and mitochondria-mediated apoptotic signaling.
TextSentencer_T106 14458-14581 Sentence denotes Our findings may provide insights into the interaction between PPV and host, and uncover the pathogenesis of PPV infection.
TextSentencer_T106 14458-14581 Sentence denotes Our findings may provide insights into the interaction between PPV and host, and uncover the pathogenesis of PPV infection.
TextSentencer_T107 14582-14712 Sentence denotes Further studies on the mechanisms of PPV-induced apoptosis should give us new insights on the interplays between PPV and its host.
TextSentencer_T107 14582-14712 Sentence denotes Further studies on the mechanisms of PPV-induced apoptosis should give us new insights on the interplays between PPV and its host.
TextSentencer_T108 14713-14955 Sentence denotes ST cells were purchased from American type culture collection (ATCC, CRL-1746) and cultured in DMEM (Gibco, US) culture medium supplemented with 10 % heat-inactivated fetal bovine serum (Gibco, US) at 37°C in a 5 % CO 2 humidified atmosphere.
TextSentencer_T108 14713-14955 Sentence denotes ST cells were purchased from American type culture collection (ATCC, CRL-1746) and cultured in DMEM (Gibco, US) culture medium supplemented with 10 % heat-inactivated fetal bovine serum (Gibco, US) at 37°C in a 5 % CO 2 humidified atmosphere.
TextSentencer_T109 14956-15013 Sentence denotes The PPV YL strain was used as previously described [19] .
TextSentencer_T109 14956-15013 Sentence denotes The PPV YL strain was used as previously described [19] .
TextSentencer_T110 15014-15135 Sentence denotes Virus titers were determined by 50 % tissue culture infective doses (TCID 50 ) according to Reed and Muench method [31] .
TextSentencer_T110 15014-15135 Sentence denotes Virus titers were determined by 50 % tissue culture infective doses (TCID 50 ) according to Reed and Muench method [31] .
TextSentencer_T111 15136-15279 Sentence denotes Mouse monoclonal antibodies against Bcl-2, Bax, Cytochrome c, Cox-4, PARP, Fas, FasL, and β-actin were purchased from Santa Cruz Biotechnology.
TextSentencer_T111 15136-15279 Sentence denotes Mouse monoclonal antibodies against Bcl-2, Bax, Cytochrome c, Cox-4, PARP, Fas, FasL, and β-actin were purchased from Santa Cruz Biotechnology.
TextSentencer_T112 15280-15378 Sentence denotes Horseradish peroxidase (HRP)-conjugated secondary antibody was purchased from Promega Corporation.
TextSentencer_T112 15280-15378 Sentence denotes Horseradish peroxidase (HRP)-conjugated secondary antibody was purchased from Promega Corporation.
TextSentencer_T113 15379-15429 Sentence denotes Caspases activities assay of caspase-3, -8, and -9
TextSentencer_T113 15379-15429 Sentence denotes Caspases activities assay of caspase-3, -8, and -9
TextSentencer_T114 15430-15530 Sentence denotes ST cells were infected with PPV at 1.0 MOI and incubated in a 5 % CO 2 humidified incubator at 37°C.
TextSentencer_T114 15430-15530 Sentence denotes ST cells were infected with PPV at 1.0 MOI and incubated in a 5 % CO 2 humidified incubator at 37°C.
TextSentencer_T115 15531-15686 Sentence denotes The cells were harvested at 0, 6, 12, 24, 36, and 48 hpi, washed with ice-cold PBS for three times, and then lysed with 200 μl lysis buffer on ice for 1 h.
TextSentencer_T115 15531-15686 Sentence denotes The cells were harvested at 0, 6, 12, 24, 36, and 48 hpi, washed with ice-cold PBS for three times, and then lysed with 200 μl lysis buffer on ice for 1 h.
TextSentencer_T116 15687-15798 Sentence denotes The caspases activities were tested with colorimetric assay kit (Abcam, US) following the manufacture's manual.
TextSentencer_T116 15687-15798 Sentence denotes The caspases activities were tested with colorimetric assay kit (Abcam, US) following the manufacture's manual.
TextSentencer_T117 15799-15892 Sentence denotes Briefly, the protein concentration was measured using BCA Protein Assay Reagent (Pierce, US).
TextSentencer_T117 15799-15892 Sentence denotes Briefly, the protein concentration was measured using BCA Protein Assay Reagent (Pierce, US).
TextSentencer_T118 15893-16006 Sentence denotes 150 μg total protein of each sample was incubated with each caspase substrate in a 96-well plate for 4 h at 37°C.
TextSentencer_T118 15893-16006 Sentence denotes 150 μg total protein of each sample was incubated with each caspase substrate in a 96-well plate for 4 h at 37°C.
TextSentencer_T119 16007-16081 Sentence denotes The absorbance was measured at 400 nm with spectrophotometer (BioTek, US).
TextSentencer_T119 16007-16081 Sentence denotes The absorbance was measured at 400 nm with spectrophotometer (BioTek, US).
TextSentencer_T120 16082-16231 Sentence denotes The cells were harvested and stained with Tetrechlorotetraethyl benzimidazol carbocyanine iodide 1 (JC-1) for 15 min at room temperature in the dark.
TextSentencer_T120 16082-16231 Sentence denotes The cells were harvested and stained with Tetrechlorotetraethyl benzimidazol carbocyanine iodide 1 (JC-1) for 15 min at room temperature in the dark.
TextSentencer_T121 16232-16362 Sentence denotes Then the Δψm was assessed using JC-1 Mitochondrial Potential Detection Kit (Biotium Inc, US) following the manufacture's protocol.
TextSentencer_T121 16232-16362 Sentence denotes Then the Δψm was assessed using JC-1 Mitochondrial Potential Detection Kit (Biotium Inc, US) following the manufacture's protocol.
TextSentencer_T122 16363-16421 Sentence denotes The fluorescence signal was measured at 550 nm and 485 nm.
TextSentencer_T122 16363-16421 Sentence denotes The fluorescence signal was measured at 550 nm and 485 nm.
TextSentencer_T123 16422-16573 Sentence denotes ST cells were harvested and then lysed on ice for 1 h with radioimmunoprecipitation assay (RIPA) buffer with 1 mM phenylmethyl sulfonylfluoride (PMSF).
TextSentencer_T123 16422-16573 Sentence denotes ST cells were harvested and then lysed on ice for 1 h with radioimmunoprecipitation assay (RIPA) buffer with 1 mM phenylmethyl sulfonylfluoride (PMSF).
TextSentencer_T124 16574-16742 Sentence denotes Isolation of mitochondrial and cytosolic proteins was carried out using the Mitochondria/cytosol Fractionation Kit (Pierce, US) according to the manufacture's protocol.
TextSentencer_T124 16574-16742 Sentence denotes Isolation of mitochondrial and cytosolic proteins was carried out using the Mitochondria/cytosol Fractionation Kit (Pierce, US) according to the manufacture's protocol.
TextSentencer_T125 16743-16823 Sentence denotes Protein concentration was detected using BCA Protein Assay Reagent (Pierce, US).
TextSentencer_T125 16743-16823 Sentence denotes Protein concentration was detected using BCA Protein Assay Reagent (Pierce, US).
TextSentencer_T126 16824-17011 Sentence denotes Equivalent amount of protein was loaded and (See figure on previous page.) Fig. 4 The role of ROS in PPV-induced apoptosis. a The effects of ROS inhibitors PDTC and NAC on ROS production.
TextSentencer_T126 16824-17011 Sentence denotes Equivalent amount of protein was loaded and (See figure on previous page.) Fig. 4 The role of ROS in PPV-induced apoptosis. a The effects of ROS inhibitors PDTC and NAC on ROS production.
TextSentencer_T127 17012-17086 Sentence denotes PPV-infected cells were treated with PDTC or NAC and stained with DCFH-DA.
TextSentencer_T127 17012-17086 Sentence denotes PPV-infected cells were treated with PDTC or NAC and stained with DCFH-DA.
TextSentencer_T128 17087-17190 Sentence denotes The ROS production was measured by flow cytometry. b The effect of ROS on Δψm in PPV-infected ST cells.
TextSentencer_T128 17087-17190 Sentence denotes The ROS production was measured by flow cytometry. b The effect of ROS on Δψm in PPV-infected ST cells.
TextSentencer_T129 17191-17380 Sentence denotes The Δψm of cells infected with PPV and treated with PDTC or NAC was detected by flow cytometry. c The effects of ROS scavengers on the Bcl-2, Bax translocation, cytochrome c redistribution.
TextSentencer_T129 17191-17380 Sentence denotes The Δψm of cells infected with PPV and treated with PDTC or NAC was detected by flow cytometry. c The effects of ROS scavengers on the Bcl-2, Bax translocation, cytochrome c redistribution.
TextSentencer_T130 17381-17471 Sentence denotes ROS inhibitors-treated cells were infected with PPV and analyzed using western blot assay.
TextSentencer_T130 17381-17471 Sentence denotes ROS inhibitors-treated cells were infected with PPV and analyzed using western blot assay.
TextSentencer_T131 17472-17653 Sentence denotes The mitochondrial and the cytosolic proteins were respectively normalized to Cox-4 and β-actin. d The effect of ROS scavengers on the activity of caspase-3 in PPV-infected ST cells.
TextSentencer_T131 17472-17653 Sentence denotes The mitochondrial and the cytosolic proteins were respectively normalized to Cox-4 and β-actin. d The effect of ROS scavengers on the activity of caspase-3 in PPV-infected ST cells.
TextSentencer_T132 17654-17793 Sentence denotes Caspase-3 activity was measured using the caspase activity kit. e The effect of ROS scavengers on the apoptotic rate of PPV-infected cells.
TextSentencer_T132 17654-17793 Sentence denotes Caspase-3 activity was measured using the caspase activity kit. e The effect of ROS scavengers on the apoptotic rate of PPV-infected cells.
TextSentencer_T133 17794-17844 Sentence denotes The apoptotic rate was measured by flow cytometry.
TextSentencer_T133 17794-17844 Sentence denotes The apoptotic rate was measured by flow cytometry.
TextSentencer_T134 17845-18058 Sentence denotes The data represent three independent experiments ± SD. **, p < 0.01 in comparison with PPV infection alone subjected to electrophoresis on 12 % sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
TextSentencer_T134 17845-18058 Sentence denotes The data represent three independent experiments ± SD. **, p < 0.01 in comparison with PPV infection alone subjected to electrophoresis on 12 % sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
TextSentencer_T135 18059-18166 Sentence denotes Subsequently, proteins were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore Corp, US).
TextSentencer_T135 18059-18166 Sentence denotes Subsequently, proteins were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore Corp, US).
TextSentencer_T136 18167-18384 Sentence denotes The membranes were blocked with 5 % non-fat dry milk for 1 h at room temperature, and incubated with primary antibody at the dilution of 1:500 in antibody dilution buffer (PBS, 2 % non-fat dry milk) over night at 4°C.
TextSentencer_T136 18167-18384 Sentence denotes The membranes were blocked with 5 % non-fat dry milk for 1 h at room temperature, and incubated with primary antibody at the dilution of 1:500 in antibody dilution buffer (PBS, 2 % non-fat dry milk) over night at 4°C.
TextSentencer_T137 18385-18586 Sentence denotes The blots were subjected to secondary antibody at the dilution of 1:10000 in antibody dilution buffer (PBS, 2 % non-fat dry milk) at room temperature for 1 h and detected with ECL reagent (Pierce, US).
TextSentencer_T137 18385-18586 Sentence denotes The blots were subjected to secondary antibody at the dilution of 1:10000 in antibody dilution buffer (PBS, 2 % non-fat dry milk) at room temperature for 1 h and detected with ECL reagent (Pierce, US).
TextSentencer_T138 18587-18669 Sentence denotes The ROS were detected using Reactive Oxygen Species Assay Kit (Sigma-Aldrich, US).
TextSentencer_T138 18587-18669 Sentence denotes The ROS were detected using Reactive Oxygen Species Assay Kit (Sigma-Aldrich, US).
TextSentencer_T139 18670-18752 Sentence denotes Briefly, ST cells were seeded into 24-well plate and infected with PPV at 1.0 MOI.
TextSentencer_T139 18670-18752 Sentence denotes Briefly, ST cells were seeded into 24-well plate and infected with PPV at 1.0 MOI.
TextSentencer_T140 18753-18935 Sentence denotes Then the cells were stained with 10 μM 2' , 7'dichlorodihydrofluorescein (DCFH-DA) at 0, 12, 24, 36, and 48 hpi and observed under the fluorescence microscope (AMG EVOS Inc, Israel).
TextSentencer_T140 18753-18935 Sentence denotes Then the cells were stained with 10 μM 2' , 7'dichlorodihydrofluorescein (DCFH-DA) at 0, 12, 24, 36, and 48 hpi and observed under the fluorescence microscope (AMG EVOS Inc, Israel).
TextSentencer_T141 18936-19061 Sentence denotes Mock-infected and PPV-infected cells were harvested at indicated times and washed twice with DMEM without fetal bovine serum.
TextSentencer_T141 18936-19061 Sentence denotes Mock-infected and PPV-infected cells were harvested at indicated times and washed twice with DMEM without fetal bovine serum.
TextSentencer_T142 19062-19210 Sentence denotes Then the cells were incubated with 10 μM DCFH-DA at 37°C for 30 min in the dark and analyzed using flow cytometry (Beckman Coulter EPICS ALTRA, US).
TextSentencer_T142 19062-19210 Sentence denotes Then the cells were incubated with 10 μM DCFH-DA at 37°C for 30 min in the dark and analyzed using flow cytometry (Beckman Coulter EPICS ALTRA, US).
TextSentencer_T143 19211-19291 Sentence denotes Statistical analyses were calculated using one way analysis of variance (ANOVA).
TextSentencer_T143 19211-19291 Sentence denotes Statistical analyses were calculated using one way analysis of variance (ANOVA).
TextSentencer_T144 19292-19330 Sentence denotes Significance was attained at p < 0.05.
TextSentencer_T144 19292-19330 Sentence denotes Significance was attained at p < 0.05.