Id |
Subject |
Object |
Predicate |
Lexical cue |
T37 |
0-7 |
Sentence |
denotes |
Results |
T38 |
9-48 |
Sentence |
denotes |
Regulation of AP-1 and NF-κB activation |
T3051 |
49-259 |
Sentence |
denotes |
The transcription factors NF-κB and AP-1 play key roles in the initiation of an inflammatory response by inducing the expression and secretion of chemokines and cytokines that attract and activate immune cells. |
T39 |
49-259 |
Sentence |
denotes |
The transcription factors NF-κB and AP-1 play key roles in the initiation of an inflammatory response by inducing the expression and secretion of chemokines and cytokines that attract and activate immune cells. |
T3052 |
260-404 |
Sentence |
denotes |
However, the signal transduction pathways and subsequent inflammatory cytokine induction by these transcription factors is not fully elucidated. |
T40 |
260-404 |
Sentence |
denotes |
However, the signal transduction pathways and subsequent inflammatory cytokine induction by these transcription factors is not fully elucidated. |
T3053 |
405-518 |
Sentence |
denotes |
The present study is aimed at determining the involvement of AP-1 and NF-κB in cytokine induction and regulation. |
T41 |
405-518 |
Sentence |
denotes |
The present study is aimed at determining the involvement of AP-1 and NF-κB in cytokine induction and regulation. |
T3054 |
519-658 |
Sentence |
denotes |
PMA treatment resulted in an up-regulation of AP-1 after 2 h exposure and continued to increase throughout the analysis period (figure 1a). |
T42 |
519-658 |
Sentence |
denotes |
PMA treatment resulted in an up-regulation of AP-1 after 2 h exposure and continued to increase throughout the analysis period (figure 1a). |
T3055 |
659-741 |
Sentence |
denotes |
HK E. coli treatment did not affect AP-1 activation in Jurkat T-cells (figure 1b). |
T43 |
659-741 |
Sentence |
denotes |
HK E. coli treatment did not affect AP-1 activation in Jurkat T-cells (figure 1b). |
T3056 |
742-999 |
Sentence |
denotes |
To determine the involvement of associated pathways, we exposed cells to Ca2+ ionophore with or without PMA and observed a modest involvement of Ca2+ in PMA-dependent AP-1 activation (figure 1c) while Ca2+ alone did not alter AP-1 activity (data not shown). |
T44 |
742-999 |
Sentence |
denotes |
To determine the involvement of associated pathways, we exposed cells to Ca2+ ionophore with or without PMA and observed a modest involvement of Ca2+ in PMA-dependent AP-1 activation (figure 1c) while Ca2+ alone did not alter AP-1 activity (data not shown). |
T3057 |
1000-1218 |
Sentence |
denotes |
Furthermore, AP-1 activity decreased in a TCR-deficient Jurkat cell line when exposed to PMA compared to the parent cell line indicating that regulation of AP-1 was only partially T-cell receptor dependent (figure 1d). |
T45 |
1000-1218 |
Sentence |
denotes |
Furthermore, AP-1 activity decreased in a TCR-deficient Jurkat cell line when exposed to PMA compared to the parent cell line indicating that regulation of AP-1 was only partially T-cell receptor dependent (figure 1d). |
T46 |
1219-1299 |
Sentence |
denotes |
Figure 1 AP-1 activation following long-term exposure of Jurkat T-cells to PMA. |
T15023 |
1229-1299 |
Sentence |
denotes |
AP-1 activation following long-term exposure of Jurkat T-cells to PMA. |
T15024 |
1300-1727 |
Sentence |
denotes |
Jurkat T-cells were transfected with luciferase reporter plasmids containing the AP-1 cis-elements. (A) Time-dependent AP-1 activation in response to PMA. (B) HK E. coli does not activate AP-1. (C) Dose-dependent activation of AP-1 were performed using PMA alone (grey bars) and in combination with calcium ionophore (CaI 955 μM, black bars). (D) TCR-/- deficient cells responded to PMA (162 nM) by up-regulating AP-1 activity. |
T47 |
1300-1727 |
Sentence |
denotes |
Jurkat T-cells were transfected with luciferase reporter plasmids containing the AP-1 cis-elements. (A) Time-dependent AP-1 activation in response to PMA. (B) HK E. coli does not activate AP-1. (C) Dose-dependent activation of AP-1 were performed using PMA alone (grey bars) and in combination with calcium ionophore (CaI 955 μM, black bars). (D) TCR-/- deficient cells responded to PMA (162 nM) by up-regulating AP-1 activity. |
T15025 |
1728-1817 |
Sentence |
denotes |
Statistical significance from the control was determined using Student's t-test. (n = 4). |
T48 |
1728-1817 |
Sentence |
denotes |
Statistical significance from the control was determined using Student's t-test. (n = 4). |
T15026 |
1818-1853 |
Sentence |
denotes |
Controls were arbitrarily set to 1. |
T49 |
1818-1853 |
Sentence |
denotes |
Controls were arbitrarily set to 1. |
T3058 |
1856-1940 |
Sentence |
denotes |
NF-κB levels showed a transient increase at 1 min after exposure to PMA (figure 2a). |
T50 |
1856-1940 |
Sentence |
denotes |
NF-κB levels showed a transient increase at 1 min after exposure to PMA (figure 2a). |
T3059 |
1941-2076 |
Sentence |
denotes |
However, 1 h after exposure the NF-κB levels began to drop reaching the lowest levels by 6 h, after which they increased again by 24 h. |
T51 |
1941-2076 |
Sentence |
denotes |
However, 1 h after exposure the NF-κB levels began to drop reaching the lowest levels by 6 h, after which they increased again by 24 h. |
T3060 |
2077-2258 |
Sentence |
denotes |
Exposure of Jurkat T-cells to HK E. coli resulted in a dose-dependent NF-κB activation, with the highest activity observed at a relative concentration of 5 × 107 CFU/ml (figure 2b). |
T52 |
2077-2258 |
Sentence |
denotes |
Exposure of Jurkat T-cells to HK E. coli resulted in a dose-dependent NF-κB activation, with the highest activity observed at a relative concentration of 5 × 107 CFU/ml (figure 2b). |
T3061 |
2259-2468 |
Sentence |
denotes |
The time-dependent activation of NF-κB by HK E. coli was assessed further using the optimal concentration obtained from figure 2b and showed that the NF-κB activity increased after 3 h of exposure (figure 2c). |
T53 |
2259-2468 |
Sentence |
denotes |
The time-dependent activation of NF-κB by HK E. coli was assessed further using the optimal concentration obtained from figure 2b and showed that the NF-κB activity increased after 3 h of exposure (figure 2c). |
T3062 |
2469-2634 |
Sentence |
denotes |
Furthermore, increased intracellular Ca2+ reversed the PMA dependent NF-κB inhibition (figure 2d) and reduced the HK E. coli -dependent NF-κB activation (figure 2e). |
T54 |
2469-2634 |
Sentence |
denotes |
Furthermore, increased intracellular Ca2+ reversed the PMA dependent NF-κB inhibition (figure 2d) and reduced the HK E. coli -dependent NF-κB activation (figure 2e). |
T55 |
2635-2754 |
Sentence |
denotes |
Figure 2 NF-κB down-regulation by PMA and up-regulation by heat killed E. coli MG1655 following long-term stimulation. |
T15303 |
2645-2754 |
Sentence |
denotes |
NF-κB down-regulation by PMA and up-regulation by heat killed E. coli MG1655 following long-term stimulation. |
T15304 |
2755-2925 |
Sentence |
denotes |
Transfection of Jurkat T-cells was performed using luciferase reporter plasmids containing NF-κB cis-elements. (A) Time-dependent stimulation of Jurkat T-cells using PMA. |
T56 |
2755-2925 |
Sentence |
denotes |
Transfection of Jurkat T-cells was performed using luciferase reporter plasmids containing NF-κB cis-elements. (A) Time-dependent stimulation of Jurkat T-cells using PMA. |
T15305 |
2926-3019 |
Sentence |
denotes |
NF-κB activation was evaluated using HK E. coli in a (B) dose- and (C) time-dependant manner. |
T57 |
2926-3019 |
Sentence |
denotes |
NF-κB activation was evaluated using HK E. coli in a (B) dose- and (C) time-dependant manner. |
T15306 |
3020-3170 |
Sentence |
denotes |
Calcium ionophore increased NF-κB activity following PMA exposure (E) and resulted in a negative regulation in response to HK E. coli stimulation (D). |
T58 |
3020-3170 |
Sentence |
denotes |
Calcium ionophore increased NF-κB activity following PMA exposure (E) and resulted in a negative regulation in response to HK E. coli stimulation (D). |
T15307 |
3171-3260 |
Sentence |
denotes |
Statistical significance from the control was determined using Student's t-test. (n = 4). |
T59 |
3171-3260 |
Sentence |
denotes |
Statistical significance from the control was determined using Student's t-test. (n = 4). |
T15308 |
3261-3296 |
Sentence |
denotes |
Controls were arbitrarily set to 1. |
T60 |
3261-3296 |
Sentence |
denotes |
Controls were arbitrarily set to 1. |
T3931 |
3298-3333 |
Sentence |
denotes |
Induction of inflammatory responses |
T61 |
3298-3333 |
Sentence |
denotes |
Induction of inflammatory responses |
T3932 |
3334-3495 |
Sentence |
denotes |
The ability of PMA and HK E. coli to induce an inflammatory response in Jurkat T-cells was evaluated using a multiplex cytokine assay following 24 h stimulation. |
T62 |
3334-3495 |
Sentence |
denotes |
The ability of PMA and HK E. coli to induce an inflammatory response in Jurkat T-cells was evaluated using a multiplex cytokine assay following 24 h stimulation. |
T3933 |
3496-3622 |
Sentence |
denotes |
The cytokine profile revealed an enhanced induction of the pro-inflammatory cytokines IL-2, IL-6, TNF and the chemokine CXCL8. |
T63 |
3496-3622 |
Sentence |
denotes |
The cytokine profile revealed an enhanced induction of the pro-inflammatory cytokines IL-2, IL-6, TNF and the chemokine CXCL8. |
T3934 |
3623-3754 |
Sentence |
denotes |
The levels of the anti-inflammatory cytokine IL-10 were unaffected by PMA but were significantly decreased by HK E. coli (Table 1). |
T64 |
3623-3754 |
Sentence |
denotes |
The levels of the anti-inflammatory cytokine IL-10 were unaffected by PMA but were significantly decreased by HK E. coli (Table 1). |
T3935 |
3755-3858 |
Sentence |
denotes |
These results confirmed that PMA and HK E. coli induced an inflammatory response in the Jurkat T-cells. |
T65 |
3755-3858 |
Sentence |
denotes |
These results confirmed that PMA and HK E. coli induced an inflammatory response in the Jurkat T-cells. |
T3936 |
3859-3956 |
Sentence |
denotes |
It is interesting to note that PMA was 120-fold more effective at inducing CXCL8 than HK E. coli. |
T66 |
3859-3956 |
Sentence |
denotes |
It is interesting to note that PMA was 120-fold more effective at inducing CXCL8 than HK E. coli. |
T3937 |
3957-4070 |
Sentence |
denotes |
PMA-dependent induction of AP-1 and down-regulation of NF-κB suggests an involvement of AP-1 in CXCL8 regulation. |
T67 |
3957-4070 |
Sentence |
denotes |
PMA-dependent induction of AP-1 and down-regulation of NF-κB suggests an involvement of AP-1 in CXCL8 regulation. |
T3938 |
4071-4258 |
Sentence |
denotes |
Determination of the time course of cytokine induction in response to PMA showed that CXCL8 was already released between 2-6 h, while TNF and IL-6 were released between 6-24 h (figure 3). |
T68 |
4071-4258 |
Sentence |
denotes |
Determination of the time course of cytokine induction in response to PMA showed that CXCL8 was already released between 2-6 h, while TNF and IL-6 were released between 6-24 h (figure 3). |
T3939 |
4259-4416 |
Sentence |
denotes |
These results indicated that the cytokines were differentially regulated and that the release was not associated with the early transient induction of NF-κB. |
T69 |
4259-4416 |
Sentence |
denotes |
These results indicated that the cytokines were differentially regulated and that the release was not associated with the early transient induction of NF-κB. |
T3940 |
4417-4517 |
Sentence |
denotes |
The temporal induction of AP-1 correlated to the CXCL8 levels and preceded the TNF and IL-6 release. |
T70 |
4417-4517 |
Sentence |
denotes |
The temporal induction of AP-1 correlated to the CXCL8 levels and preceded the TNF and IL-6 release. |
T3941 |
4518-4589 |
Sentence |
denotes |
This suggests an association between CXCL8 release and AP-1 signalling. |
T71 |
4518-4589 |
Sentence |
denotes |
This suggests an association between CXCL8 release and AP-1 signalling. |
T72 |
4590-4672 |
Sentence |
denotes |
Figure 3 CXCL8, IL-6 and TNF expression following long-term stimulation with PMA. |
T15595 |
4600-4672 |
Sentence |
denotes |
CXCL8, IL-6 and TNF expression following long-term stimulation with PMA. |
T15596 |
4673-4809 |
Sentence |
denotes |
Jurkat T-cells were either treated with media (white bars) or stimulated with PMA (black bars) and incubated for 1 h, 2 h, 6 h and 24 h. |
T73 |
4673-4809 |
Sentence |
denotes |
Jurkat T-cells were either treated with media (white bars) or stimulated with PMA (black bars) and incubated for 1 h, 2 h, 6 h and 24 h. |
T15597 |
4810-4933 |
Sentence |
denotes |
Aged media was added following each centrifugation step representative to the stimulation time (see Materials and methods). |
T74 |
4810-4933 |
Sentence |
denotes |
Aged media was added following each centrifugation step representative to the stimulation time (see Materials and methods). |
T15598 |
4934-5005 |
Sentence |
denotes |
Cytokine levels (A) CXCL8, (B) IL-6 and (C) TNF were detected by ELISA. |
T75 |
4934-5005 |
Sentence |
denotes |
Cytokine levels (A) CXCL8, (B) IL-6 and (C) TNF were detected by ELISA. |
T15599 |
5006-5095 |
Sentence |
denotes |
Statistical significance from the control was determined using Student's t-test. (n = 3). |
T76 |
5006-5095 |
Sentence |
denotes |
Statistical significance from the control was determined using Student's t-test. (n = 3). |
T77 |
5096-5191 |
Sentence |
denotes |
Table 1 Jurkat T-cells were stimulated with 162 nM PMA and 5 × 107 CFU/ml HK E. coli for 24 h. |
T17005 |
5105-5191 |
Sentence |
denotes |
Jurkat T-cells were stimulated with 162 nM PMA and 5 × 107 CFU/ml HK E. coli for 24 h. |
T78 |
5193-5248 |
Sentence |
denotes |
Cytokine levels were determined using multiplex assays. |
T79 |
5249-5323 |
Sentence |
denotes |
Concentrations are given as pg/ml for each group (mean value ± SD, n = 3). |
T80 |
5324-5408 |
Sentence |
denotes |
Statistical significance from the control (C) was determined using Student's t-test. |
T4753 |
5410-5462 |
Sentence |
denotes |
Cooperative induction of cytokines by AP-1 and NF-κB |
T81 |
5410-5462 |
Sentence |
denotes |
Cooperative induction of cytokines by AP-1 and NF-κB |
T4754 |
5463-5594 |
Sentence |
denotes |
To further characterize the involvement of NF-κB in cytokine regulation, we treated cells with an NF-κB activation inhibitor (NAI). |
T82 |
5463-5594 |
Sentence |
denotes |
To further characterize the involvement of NF-κB in cytokine regulation, we treated cells with an NF-κB activation inhibitor (NAI). |
T4755 |
5595-5739 |
Sentence |
denotes |
The results showed that NAI selectively down-regulated NF-κB activation (figures 4a and 4b) and did not alter AP-1 activity (figures 4c and 4d). |
T83 |
5595-5739 |
Sentence |
denotes |
The results showed that NAI selectively down-regulated NF-κB activation (figures 4a and 4b) and did not alter AP-1 activity (figures 4c and 4d). |
T4756 |
5740-5921 |
Sentence |
denotes |
Exposure of Jurkat T-cells to NAI resulted in a modest reduction of CXCL8 following PMA exposure, while it did not alter the CXCL8 release following HK E. coli exposure (figure 5a). |
T84 |
5740-5921 |
Sentence |
denotes |
Exposure of Jurkat T-cells to NAI resulted in a modest reduction of CXCL8 following PMA exposure, while it did not alter the CXCL8 release following HK E. coli exposure (figure 5a). |
T4757 |
5922-6094 |
Sentence |
denotes |
NAI did not affect TNF expression (figure 5b) indicating that NF-κB is not the main regulator of CXCL8 or TNF following either PMA or HK E. coli exposure in Jurkat T-cells. |
T85 |
5922-6094 |
Sentence |
denotes |
NAI did not affect TNF expression (figure 5b) indicating that NF-κB is not the main regulator of CXCL8 or TNF following either PMA or HK E. coli exposure in Jurkat T-cells. |
T4758 |
6095-6295 |
Sentence |
denotes |
In contrast, NAI resulted in a complete inhibition of IL-6 following PMA exposure and a 45% inhibition following HK E. coli exposure (figure 5c), suggesting an involvement of NF-κB in IL-6 regulation. |
T86 |
6095-6295 |
Sentence |
denotes |
In contrast, NAI resulted in a complete inhibition of IL-6 following PMA exposure and a 45% inhibition following HK E. coli exposure (figure 5c), suggesting an involvement of NF-κB in IL-6 regulation. |
T87 |
6296-6342 |
Sentence |
denotes |
Figure 4 Inhibition of NF-κB activity by NAI. |
T15871 |
6306-6342 |
Sentence |
denotes |
Inhibition of NF-κB activity by NAI. |
T15872 |
6343-6461 |
Sentence |
denotes |
Jurkat T-cells were transfected with luciferase reporter plasmids containing either NF-κB or AP-1 cis-acting elements. |
T88 |
6343-6461 |
Sentence |
denotes |
Jurkat T-cells were transfected with luciferase reporter plasmids containing either NF-κB or AP-1 cis-acting elements. |
T15873 |
6462-6682 |
Sentence |
denotes |
The cells were incubated with NF-κB activation inhibitor (NAI) for 1 h followed by stimulation with PMA (162 nM, A and C) or HK E. coli (5 × 107 CFU/ml, B and D) for 24 h. (A, B) NF-κB, but not (C, D) AP-1 was inhibited. |
T89 |
6462-6682 |
Sentence |
denotes |
The cells were incubated with NF-κB activation inhibitor (NAI) for 1 h followed by stimulation with PMA (162 nM, A and C) or HK E. coli (5 × 107 CFU/ml, B and D) for 24 h. (A, B) NF-κB, but not (C, D) AP-1 was inhibited. |
T15874 |
6683-6798 |
Sentence |
denotes |
Statistical significance from the positive control (PMA/HK E. coli) was determined using Student's t-test. (n = 4). |
T90 |
6683-6798 |
Sentence |
denotes |
Statistical significance from the positive control (PMA/HK E. coli) was determined using Student's t-test. (n = 4). |
T15875 |
6799-6822 |
Sentence |
denotes |
Controls were set to 1. |
T91 |
6799-6822 |
Sentence |
denotes |
Controls were set to 1. |
T92 |
6823-6877 |
Sentence |
denotes |
Figure 5 Involvement of NF-κB in cytokine regulation. |
T16165 |
6833-6877 |
Sentence |
denotes |
Involvement of NF-κB in cytokine regulation. |
T16166 |
6878-7474 |
Sentence |
denotes |
Cytokine/chemokine levels were determined using ELISA following incubation of Jurkat T-cells with NAI (1 h) and stimulation (24 h). (A) CXCL8 expression was partially inhibited following PMA stimulation (grey bars), whereas the levels were not altered following stimulation with HK E. coli (black bars), this indicates an induction mainly regulated by AP-1. (B) TNF expression was not affected by NAI. (C) IL-6 release was completely inhibited by NAI following PMA exposure, indicating regulation through NF-κB since IL-6 expression was significantly increased in response to HK E. coli than PMA. |
T93 |
6878-7474 |
Sentence |
denotes |
Cytokine/chemokine levels were determined using ELISA following incubation of Jurkat T-cells with NAI (1 h) and stimulation (24 h). (A) CXCL8 expression was partially inhibited following PMA stimulation (grey bars), whereas the levels were not altered following stimulation with HK E. coli (black bars), this indicates an induction mainly regulated by AP-1. (B) TNF expression was not affected by NAI. (C) IL-6 release was completely inhibited by NAI following PMA exposure, indicating regulation through NF-κB since IL-6 expression was significantly increased in response to HK E. coli than PMA. |
T16167 |
7475-7590 |
Sentence |
denotes |
Statistical significance from the positive control (PMA/HK E. coli) was determined using Student's t-test. (n = 3). |
T94 |
7475-7590 |
Sentence |
denotes |
Statistical significance from the positive control (PMA/HK E. coli) was determined using Student's t-test. (n = 3). |
T4759 |
7593-7802 |
Sentence |
denotes |
Ca2+ was observed to increase AP-1 activity (figure 1c) and reduce NF-κB activity (figure 2e); therefore, we exposed T-cells to a PKC inhibitor together with PMA to determine its effect on cytokine expression. |
T95 |
7593-7802 |
Sentence |
denotes |
Ca2+ was observed to increase AP-1 activity (figure 1c) and reduce NF-κB activity (figure 2e); therefore, we exposed T-cells to a PKC inhibitor together with PMA to determine its effect on cytokine expression. |
T4760 |
7803-7926 |
Sentence |
denotes |
Inhibition of PKC reduced CXCL8 release from 7 ng/ml to 3 ng/ml while it had a modest effect on IL-6 and TNF (figure 6a-c). |
T96 |
7803-7926 |
Sentence |
denotes |
Inhibition of PKC reduced CXCL8 release from 7 ng/ml to 3 ng/ml while it had a modest effect on IL-6 and TNF (figure 6a-c). |
T4761 |
7927-8016 |
Sentence |
denotes |
This prompted us to test the effect of JNK inhibition on PMA-induced cytokine expression. |
T97 |
7927-8016 |
Sentence |
denotes |
This prompted us to test the effect of JNK inhibition on PMA-induced cytokine expression. |
T4762 |
8017-8207 |
Sentence |
denotes |
JNK is involved in the regulation of a multitude of different transcription factors, including the phosphorylation and activation of c-Jun, c-Fos and p53, leading to cellular apoptosis [26]. |
T98 |
8017-8207 |
Sentence |
denotes |
JNK is involved in the regulation of a multitude of different transcription factors, including the phosphorylation and activation of c-Jun, c-Fos and p53, leading to cellular apoptosis [26]. |
T4763 |
8208-8360 |
Sentence |
denotes |
Inhibition of the JNK pathways resulted in a down-regulation of both CXCL8 and IL-6, while no clear effect was observed on TNF expression (figure 6a-c). |
T99 |
8208-8360 |
Sentence |
denotes |
Inhibition of the JNK pathways resulted in a down-regulation of both CXCL8 and IL-6, while no clear effect was observed on TNF expression (figure 6a-c). |
T4764 |
8361-8497 |
Sentence |
denotes |
Analysis of mRNA levels using RT-qPCR (table 2) showed that PMA induced both il-6 and cxcl8 mRNA (5.1-fold and 111.8 fold respectively). |
T100 |
8361-8497 |
Sentence |
denotes |
Analysis of mRNA levels using RT-qPCR (table 2) showed that PMA induced both il-6 and cxcl8 mRNA (5.1-fold and 111.8 fold respectively). |
T4765 |
8498-8603 |
Sentence |
denotes |
Addition of the NF-κB inhibitor NAI and the JNK inhibitor reduced the il-6 expression below basal levels. |
T101 |
8498-8603 |
Sentence |
denotes |
Addition of the NF-κB inhibitor NAI and the JNK inhibitor reduced the il-6 expression below basal levels. |
T4766 |
8604-8726 |
Sentence |
denotes |
In contrast, while the cxcl8 levels were suppressed by the same treatments the levels remained elevated above basal level. |
T102 |
8604-8726 |
Sentence |
denotes |
In contrast, while the cxcl8 levels were suppressed by the same treatments the levels remained elevated above basal level. |
T103 |
8727-8787 |
Sentence |
denotes |
Figure 6 Involvement of PKC and JNK in cytokine expression. |
T16436 |
8737-8787 |
Sentence |
denotes |
Involvement of PKC and JNK in cytokine expression. |
T16437 |
8788-9128 |
Sentence |
denotes |
Jurkat T-cells were incubated with PKC- and JNK inhibitors for 1 h followed by stimulation with PMA (162 nM) for 24 h. (A) CXCL8 expression was inhibited in a dose-dependent manner by both inhibitors. (B) JNK-I revealed a stronger inhibitory effect on IL-6 expression than the PKC-I. (C) TNF expression was not affected by either inhibitor. |
T104 |
8788-9128 |
Sentence |
denotes |
Jurkat T-cells were incubated with PKC- and JNK inhibitors for 1 h followed by stimulation with PMA (162 nM) for 24 h. (A) CXCL8 expression was inhibited in a dose-dependent manner by both inhibitors. (B) JNK-I revealed a stronger inhibitory effect on IL-6 expression than the PKC-I. (C) TNF expression was not affected by either inhibitor. |
T16438 |
9129-9233 |
Sentence |
denotes |
Statistical significance from the positive control (PMA) was determined using Student's t-test. (n = 3). |
T105 |
9129-9233 |
Sentence |
denotes |
Statistical significance from the positive control (PMA) was determined using Student's t-test. (n = 3). |
T106 |
9234-9368 |
Sentence |
denotes |
Table 2 Jurkat T-cells were treated with 10 nM NAI, 10 μM JNK-I or10 nM PKC-I for 2 h followed by induction with 162 nM PMA for 24 h. |
T17046 |
9243-9368 |
Sentence |
denotes |
Jurkat T-cells were treated with 10 nM NAI, 10 μM JNK-I or10 nM PKC-I for 2 h followed by induction with 162 nM PMA for 24 h. |
T107 |
9370-9415 |
Sentence |
denotes |
Gene expression was determined using RT-qPCR. |
T108 |
9416-9458 |
Sentence |
denotes |
The Ct values were normalized against 18S. |
T109 |
9459-9554 |
Sentence |
denotes |
Statistical significance from the positive control (PMA) was determined using Student's t-test. |
T110 |
9555-9632 |
Sentence |
denotes |
Concentrations are given as fg/ml, values are presented as mean ± SEM, n = 3. |
T5994 |
9634-9689 |
Sentence |
denotes |
NF-κB inhibition due to PKC-dependent Bcl10 degradation |
T111 |
9634-9689 |
Sentence |
denotes |
NF-κB inhibition due to PKC-dependent Bcl10 degradation |
T5995 |
9690-9885 |
Sentence |
denotes |
Western blot analysis revealed an up-regulation of phosphorylated-PKC after 24 h treatment of Jurkat T-cells with PMA or HK E. coli (figure 7a), while IκBβ levels remained unaffected (figure 7b). |
T112 |
9690-9885 |
Sentence |
denotes |
Western blot analysis revealed an up-regulation of phosphorylated-PKC after 24 h treatment of Jurkat T-cells with PMA or HK E. coli (figure 7a), while IκBβ levels remained unaffected (figure 7b). |
T5996 |
9886-10052 |
Sentence |
denotes |
Bcl10 is a signalling protein that acts upstream of NF-κB in concert with CARMA1 and MALT1 and has been suggested to directly regulate NF-κB activity in T-cells [27]. |
T113 |
9886-10052 |
Sentence |
denotes |
Bcl10 is a signalling protein that acts upstream of NF-κB in concert with CARMA1 and MALT1 and has been suggested to directly regulate NF-κB activity in T-cells [27]. |
T5997 |
10053-10197 |
Sentence |
denotes |
Therefore, Bcl10 activation was evaluated in both control and PMA stimulated cells after 10 min, 1 h, 6 h, and 24 h using western blot analysis. |
T114 |
10053-10197 |
Sentence |
denotes |
Therefore, Bcl10 activation was evaluated in both control and PMA stimulated cells after 10 min, 1 h, 6 h, and 24 h using western blot analysis. |
T5998 |
10198-10331 |
Sentence |
denotes |
The Bcl10 levels decreased following treatment with PMA, while in control cells, Bcl10 returned to higher levels by 24 h (figure 7c). |
T115 |
10198-10331 |
Sentence |
denotes |
The Bcl10 levels decreased following treatment with PMA, while in control cells, Bcl10 returned to higher levels by 24 h (figure 7c). |
T5999 |
10332-10421 |
Sentence |
denotes |
This suggests that Bcl10 is involved in the PMA dependent inhibition of NF-κB activation. |
T116 |
10332-10421 |
Sentence |
denotes |
This suggests that Bcl10 is involved in the PMA dependent inhibition of NF-κB activation. |
T117 |
10422-10502 |
Sentence |
denotes |
Figure 7 NF-κB inhibition by PMA correlated to PKC-dependent Bcl10 degradation. |
T16732 |
10432-10502 |
Sentence |
denotes |
NF-κB inhibition by PMA correlated to PKC-dependent Bcl10 degradation. |
T16733 |
10503-10973 |
Sentence |
denotes |
Levels of intracellular protein were assessed following 24 h stimulation with PMA (162 nM) or HK E. coli (5 × 107 CFU/ml). (A) Phospho-PKC increased in response to PMA and HK E. coli stimulation. (B) IκB decreased following stimulation with HK E. coli indicating NF-κB activation. (C) Bcl10 activation was inhibited following long-term stimulation with PMA, which explains the inhibitory effect of PMA on NF-κB activation. β-actin was used as a loading control. (n = 3). |
T118 |
10503-10973 |
Sentence |
denotes |
Levels of intracellular protein were assessed following 24 h stimulation with PMA (162 nM) or HK E. coli (5 × 107 CFU/ml). (A) Phospho-PKC increased in response to PMA and HK E. coli stimulation. (B) IκB decreased following stimulation with HK E. coli indicating NF-κB activation. (C) Bcl10 activation was inhibited following long-term stimulation with PMA, which explains the inhibitory effect of PMA on NF-κB activation. β-actin was used as a loading control. (n = 3). |