PMC:2674207 / 25824-28252 JSONTXT

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    2_test

    {"project":"2_test","denotations":[{"id":"19436703-17314103-86274496","span":{"begin":97,"end":99},"obj":"17314103"},{"id":"T64889","span":{"begin":97,"end":99},"obj":"17314103"}],"text":"Ligand-Activated GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    pmc-enju-pas

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GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

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GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    GO-BP

    {"project":"GO-BP","denotations":[{"id":"T9278","span":{"begin":176,"end":190},"obj":"http://purl.obolibrary.org/obo/GO_0051170"},{"id":"T9279","span":{"begin":744,"end":758},"obj":"http://purl.obolibrary.org/obo/GO_0051170"},{"id":"T17414","span":{"begin":988,"end":1002},"obj":"http://purl.obolibrary.org/obo/GO_0051170"},{"id":"T17415","span":{"begin":2214,"end":2233},"obj":"http://purl.obolibrary.org/obo/GO_0002672"},{"id":"T17416","span":{"begin":2214,"end":2233},"obj":"http://purl.obolibrary.org/obo/GO_0045579"},{"id":"T17417","span":{"begin":2214,"end":2233},"obj":"http://purl.obolibrary.org/obo/GO_0002869"},{"id":"T17418","span":{"begin":2214,"end":2233},"obj":"http://purl.obolibrary.org/obo/GO_0002904"},{"id":"T17419","span":{"begin":2214,"end":2233},"obj":"http://purl.obolibrary.org/obo/GO_0050871"},{"id":"T17420","span":{"begin":2214,"end":2233},"obj":"http://purl.obolibrary.org/obo/GO_0031296"},{"id":"T17421","span":{"begin":2214,"end":2233},"obj":"http://purl.obolibrary.org/obo/GO_0030890"}],"text":"Ligand-Activated GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    GO-MF

    {"project":"GO-MF","denotations":[{"id":"T9280","span":{"begin":0,"end":6},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T9281","span":{"begin":114,"end":120},"obj":"http://purl.obolibrary.org/obo/GO_0005488"}],"text":"Ligand-Activated GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    GO-CC

    {"project":"GO-CC","denotations":[{"id":"T17422","span":{"begin":1811,"end":1816},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T17423","span":{"begin":2163,"end":2168},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T17424","span":{"begin":2214,"end":2219},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T17425","span":{"begin":2317,"end":2322},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T18127","span":{"begin":2163,"end":2168},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"Ligand-Activated GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    sentences

    {"project":"sentences","denotations":[{"id":"T8940","span":{"begin":0,"end":55},"obj":"Sentence"},{"id":"T8941","span":{"begin":56,"end":210},"obj":"Sentence"},{"id":"T8942","span":{"begin":211,"end":337},"obj":"Sentence"},{"id":"T8943","span":{"begin":338,"end":450},"obj":"Sentence"},{"id":"T8944","span":{"begin":451,"end":652},"obj":"Sentence"},{"id":"T8945","span":{"begin":653,"end":866},"obj":"Sentence"},{"id":"T16815","span":{"begin":911,"end":1003},"obj":"Sentence"},{"id":"T16816","span":{"begin":1004,"end":1190},"obj":"Sentence"},{"id":"T16817","span":{"begin":1191,"end":1252},"obj":"Sentence"},{"id":"T16818","span":{"begin":1253,"end":1308},"obj":"Sentence"},{"id":"T16819","span":{"begin":1309,"end":1614},"obj":"Sentence"},{"id":"T16820","span":{"begin":1615,"end":1670},"obj":"Sentence"},{"id":"T16821","span":{"begin":1671,"end":1957},"obj":"Sentence"},{"id":"T16822","span":{"begin":1958,"end":2013},"obj":"Sentence"},{"id":"T16823","span":{"begin":2014,"end":2279},"obj":"Sentence"},{"id":"T16824","span":{"begin":2280,"end":2357},"obj":"Sentence"},{"id":"T16825","span":{"begin":2358,"end":2425},"obj":"Sentence"},{"id":"T183","span":{"begin":0,"end":55},"obj":"Sentence"},{"id":"T184","span":{"begin":56,"end":210},"obj":"Sentence"},{"id":"T185","span":{"begin":211,"end":337},"obj":"Sentence"},{"id":"T186","span":{"begin":338,"end":450},"obj":"Sentence"},{"id":"T187","span":{"begin":451,"end":652},"obj":"Sentence"},{"id":"T188","span":{"begin":653,"end":866},"obj":"Sentence"},{"id":"T189","span":{"begin":867,"end":1003},"obj":"Sentence"},{"id":"T190","span":{"begin":1004,"end":1190},"obj":"Sentence"},{"id":"T191","span":{"begin":1191,"end":1252},"obj":"Sentence"},{"id":"T192","span":{"begin":1253,"end":1308},"obj":"Sentence"},{"id":"T193","span":{"begin":1309,"end":1614},"obj":"Sentence"},{"id":"T194","span":{"begin":1615,"end":1670},"obj":"Sentence"},{"id":"T195","span":{"begin":1671,"end":1957},"obj":"Sentence"},{"id":"T196","span":{"begin":1958,"end":2013},"obj":"Sentence"},{"id":"T197","span":{"begin":2014,"end":2279},"obj":"Sentence"},{"id":"T198","span":{"begin":2280,"end":2357},"obj":"Sentence"},{"id":"T199","span":{"begin":2358,"end":2425},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Ligand-Activated GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    events-check-again

    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bj":"T17497"},{"id":"R15017","pred":"themeOf","subj":"T17497","obj":"T17493"},{"id":"R15018","pred":"themeOf","subj":"T17499","obj":"T17498"},{"id":"R15019","pred":"themeOf","subj":"T17500","obj":"T17501"},{"id":"R15020","pred":"themeOf","subj":"T17501","obj":"T17499"},{"id":"R15021","pred":"themeOf","subj":"T17502","obj":"T17503"},{"id":"R8059","pred":"themeOf","subj":"T9361","obj":"T9362"},{"id":"R8060","pred":"themeOf","subj":"T9364","obj":"T9363"},{"id":"R8061","pred":"themeOf","subj":"T9364","obj":"T9370"},{"id":"R8062","pred":"themeOf","subj":"T9365","obj":"T9369"},{"id":"R8063","pred":"locationOf","subj":"T9368","obj":"T9370"},{"id":"R8064","pred":"locationOf","subj":"T9368","obj":"T9369"},{"id":"R8065","pred":"themeOf","subj":"T9369","obj":"T9366"},{"id":"R8066","pred":"themeOf","subj":"T9370","obj":"T9367"},{"id":"R8067","pred":"themeOf","subj":"T9372","obj":"T9374"}],"text":"Ligand-Activated GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    bionlp-st-ge-2016-reference-tees

    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GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    bionlp-st-ge-2016-reference

    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GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    bionlp-st-ge-2016-uniprot

    {"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T9089","span":{"begin":17,"end":19},"obj":"P04150"},{"id":"T9090","span":{"begin":34,"end":40},"obj":"P23771"},{"id":"T9091","span":{"begin":131,"end":133},"obj":"P04150"},{"id":"T9092","span":{"begin":145,"end":151},"obj":"P23771"},{"id":"T9093","span":{"begin":236,"end":238},"obj":"P04150"},{"id":"T9094","span":{"begin":349,"end":351},"obj":"P04150"},{"id":"T9095","span":{"begin":545,"end":551},"obj":"P23771"},{"id":"T9096","span":{"begin":583,"end":586},"obj":"P09693"},{"id":"T9097","span":{"begin":583,"end":586},"obj":"P20963"},{"id":"T9098","span":{"begin":583,"end":586},"obj":"P07766"},{"id":"T9099","span":{"begin":583,"end":586},"obj":"P04234"},{"id":"T9100","span":{"begin":587,"end":591},"obj":"P10747"},{"id":"T9101","span":{"begin":672,"end":675},"obj":"Q9U6Y4"},{"id":"T9102","span":{"begin":685,"end":691},"obj":"P23771"},{"id":"T9103","span":{"begin":737,"end":743},"obj":"P23771"},{"id":"T9104","span":{"begin":774,"end":777},"obj":"P09693"},{"id":"T9105","span":{"begin":774,"end":777},"obj":"P07766"},{"id":"T9106","span":{"begin":774,"end":777},"obj":"P04234"},{"id":"T9107","span":{"begin":774,"end":777},"obj":"P20963"},{"id":"T9108","span":{"begin":778,"end":782},"obj":"P10747"},{"id":"T17086","span":{"begin":942,"end":948},"obj":"P23771"},{"id":"T17087","span":{"begin":981,"end":987},"obj":"P23771"},{"id":"T17088","span":{"begin":1151,"end":1153},"obj":"P04150"},{"id":"T17089","span":{"begin":1214,"end":1216},"obj":"P04150"},{"id":"T17090","span":{"begin":1574,"end":1576},"obj":"P04150"},{"id":"T17091","span":{"begin":1842,"end":1845},"obj":"P04234"},{"id":"T17092","span":{"begin":1842,"end":1845},"obj":"P07766"},{"id":"T17093","span":{"begin":1842,"end":1845},"obj":"P20963"},{"id":"T17094","span":{"begin":1842,"end":1845},"obj":"P09693"},{"id":"T17095","span":{"begin":1846,"end":1850},"obj":"P10747"},{"id":"T17096","span":{"begin":1917,"end":1923},"obj":"P23771"},{"id":"T17097","span":{"begin":2147,"end":2151},"obj":"P10747"},{"id":"T17098","span":{"begin":2174,"end":2177},"obj":"Q9U6Y4"},{"id":"T17099","span":{"begin":2185,"end":2191},"obj":"P23771"},{"id":"T17100","span":{"begin":2270,"end":2273},"obj":"P07766"},{"id":"T17101","span":{"begin":2270,"end":2273},"obj":"P20963"},{"id":"T17102","span":{"begin":2270,"end":2273},"obj":"P04234"},{"id":"T17103","span":{"begin":2270,"end":2273},"obj":"P09693"},{"id":"T17104","span":{"begin":2274,"end":2278},"obj":"P10747"},{"id":"T17850","span":{"begin":2147,"end":2151},"obj":"P10747"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"Ligand-Activated GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}

    test2

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GR Competes with GATA-3 for Importin-α\nWe confirmed and extended previous data [20] to show that ligand-activated GR as well as GATA-3 uses importin-α for its nuclear import (Figure 2A and 2B). This interaction between GR and importin-α was significant at concentrations as low as 10−12 M and was maximal with 10−8 M FP. Subsequent GR nuclear translocation was rapid and sustained at significant levels for at least 14 h (Figure 2B). Using IP-Western blotting we showed that FP at 10−12–10−8 M decreased the association between GATA-3 and importin-α induced by anti-CD3/CD28 stimulation in a concentration-dependent manner (Figure 2C). In addition, using GFP-labelled GATA-3 and confocal microscopy we demonstrated that GATA-3 nuclear import following anti-CD3/CD28 stimulation for 30 min was attenuated by pretreatment with FP (10−8 M) (Figure 2D).\n10.1371/journal.pmed.1000076.g002 Figure 2 Fluticasone propionate reduces GATA-3 association with importin-α and GATA-3 nuclear import.\n(A) Western blot analysis demonstrates a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR interaction with importin-α (Imp-α). A positive control for GR association with importin is shown. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. *** p\u003c0.001 compared to control, ### p\u003c0.001. (B) Western blot analysis demonstrated a time- (at 10−8 M FP) and concentration- (at 60 min after stimulation) dependent induction of FP-activated GR nuclear translocation measured by IP. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ***p\u003c0.001 compared to control. (C) Western blot analysis of HuT-78 cells treated with FP and anti-CD3/CD28 co-stimulation demonstrated a concentration-dependent decrease in GATA-3–importin-α association at 20 min. Quantification of the densitometry data is shown below. Each bar represents mean±SEM of at least three independent experiments. ### p\u003c0.001 compared to control, ***p\u003c0.001 compared to αCD3/CD28-stimulated cells. (D) GFP-tagged GATA-3 was overexpressed and cells stimulated (b, c) or not (a) for 30 min with anti-CD3/CD28. The effect of 30 min pretreatment of cells with FP (10−8 M, c) is also shown. All data were analysed by ANOVA followed by Newman-Keuls post-test.\n\nE"}