pathscan phospho p38 mapk thr180 tyr182 sandwich elisa  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pathscan phospho p38 mapk thr180 tyr182 sandwich elisa
    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, <t>the</t> <t>p38-MAPK</t> inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases <t>in</t> <t>phospho-Thr180/Tyr182-p38</t> were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.
    Pathscan Phospho P38 Mapk Thr180 Tyr182 Sandwich Elisa, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pathscan phospho p38 mapk thr180 tyr182 sandwich elisa/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pathscan phospho p38 mapk thr180 tyr182 sandwich elisa - by Bioz Stars, 2023-02
    86/100 stars

    Images

    1) Product Images from "SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1"

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    Journal: medRxiv

    doi: 10.1101/2023.01.24.23284890

    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.
    Figure Legend Snippet: a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.

    Techniques Used: Expressing, Activity Assay, Activation Assay, Two Tailed Test, Standard Deviation

    a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.
    Figure Legend Snippet: a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.

    Techniques Used: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Inhibition, Chromatin Immunoprecipitation, Agarose Gel Electrophoresis, Negative Control, Two Tailed Test, Standard Deviation

    A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.
    Figure Legend Snippet: A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.

    Techniques Used: Expressing, Two Tailed Test, Activity Assay

    Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]
    Figure Legend Snippet: Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]

    Techniques Used: Protein Binding, Activation Assay, Expressing, Binding Assay

    phospho p38 mapk  (Cell Signaling Technology Inc)


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    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Structured Review

    Cell Signaling Technology Inc phospho p38 mapk
    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, <t>the</t> <t>p38-MAPK</t> inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.
    Phospho P38 Mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho p38 mapk/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 mapk - by Bioz Stars, 2023-02
    86/100 stars

    Images

    1) Product Images from "SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1"

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    Journal: medRxiv

    doi: 10.1101/2023.01.24.23284890

    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.
    Figure Legend Snippet: a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.

    Techniques Used: Expressing, Activity Assay, Activation Assay, Two Tailed Test, Standard Deviation

    a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.
    Figure Legend Snippet: a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.

    Techniques Used: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Inhibition, Chromatin Immunoprecipitation, Agarose Gel Electrophoresis, Negative Control, Two Tailed Test, Standard Deviation

    A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.
    Figure Legend Snippet: A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.

    Techniques Used: Expressing, Two Tailed Test, Activity Assay

    Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]
    Figure Legend Snippet: Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]

    Techniques Used: Protein Binding, Activation Assay, Expressing, Binding Assay

    anti phospho p38 p p38  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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  • 86

    Structured Review

    Cell Signaling Technology Inc anti phospho p38 p p38
    The mechanism how interleukin‐38 suppresses macrophage activation. (a) Representative western blot analysis of <t>p‐p38,</t> p38, p‐JNK, and JNK in the aortic wall on day 14. (b) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p = 0.0280 [ t test], p = 0.0411 [Mann–Whitney U test]) and p‐JNK ( n = 6, p = 0.6893 [ t test], p = 0.6991 [Mann–Whitney U test]). (c) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in macrophages after TNF‐α stimulation with or without IL‐38. (d) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p < 0.0001 [ANOVA], p < 0.0001 [post hoc, Control vs. TNF‐α], p = 0.0234 [post hoc, Control vs. TNF‐α + IL‐38], p = 0.0125 [post hoc, TNF‐α vs. TNF‐α + IL‐38]) and p‐JNK ( n = 6, p = 0.0283 [ANOVA], p = 0.0264 [post hoc, Control vs. TNF‐α]). (e) Representative western blot analysis of iNOS, MMP‐9, MMP‐2, and GAPDH in macrophages after TNF‐α stimulation with or without IL‐38 in the presence of SB203580. (f) Bar graphs showing the relative densitometry of iNOS ( n = 3, p = 0.2000 [Mann–Whitney U test]), MMP‐9 ( n = 3, p = 0.7000 [Mann–Whitney U test]), and MMP‐2 ( n = 3, p = 0.7000 [Mann–Whitney U test]). (g) Representative photomicrographs of the aortic wall on day 14 in the presence of SB203580, stained with EVG (upper row) or anti‐CD68 antibody (lower row). (h) Bar graph showing the number of CD68 + cells in the aortic wall on day 14 in the presence of SB203580 ( n = 5–6, p = 0.6623 [Mann–Whitney U test]). (i) Bar graphs showing the aorta diameter ( n = 5–6, p = 0.2468 [Mann–Whitney U test]), AAA incidence ( n = 5–6, p = 0.2506 [χ 2 test]), adventitia ( n = 5–6, p > 0.9999 [Mann–Whitney U test]), intramedia ( n = 5–6, p = 0.1710 [Mann–Whitney U test]), and elastin degradation score ( n = 5–6, p = 0.4917 [ t test], p = 0.6753 [Mann–Whitney U test]) on day 14 in the presence of SB203580.
    Anti Phospho P38 P P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Interleukin‐38 suppresses abdominal aortic aneurysm formation in mice by regulating macrophages in an IL1RL2 ‐p38 pathway‐dependent manner"

    Article Title: Interleukin‐38 suppresses abdominal aortic aneurysm formation in mice by regulating macrophages in an IL1RL2 ‐p38 pathway‐dependent manner

    Journal: Physiological Reports

    doi: 10.14814/phy2.15581

    The mechanism how interleukin‐38 suppresses macrophage activation. (a) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in the aortic wall on day 14. (b) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p = 0.0280 [ t test], p = 0.0411 [Mann–Whitney U test]) and p‐JNK ( n = 6, p = 0.6893 [ t test], p = 0.6991 [Mann–Whitney U test]). (c) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in macrophages after TNF‐α stimulation with or without IL‐38. (d) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p < 0.0001 [ANOVA], p < 0.0001 [post hoc, Control vs. TNF‐α], p = 0.0234 [post hoc, Control vs. TNF‐α + IL‐38], p = 0.0125 [post hoc, TNF‐α vs. TNF‐α + IL‐38]) and p‐JNK ( n = 6, p = 0.0283 [ANOVA], p = 0.0264 [post hoc, Control vs. TNF‐α]). (e) Representative western blot analysis of iNOS, MMP‐9, MMP‐2, and GAPDH in macrophages after TNF‐α stimulation with or without IL‐38 in the presence of SB203580. (f) Bar graphs showing the relative densitometry of iNOS ( n = 3, p = 0.2000 [Mann–Whitney U test]), MMP‐9 ( n = 3, p = 0.7000 [Mann–Whitney U test]), and MMP‐2 ( n = 3, p = 0.7000 [Mann–Whitney U test]). (g) Representative photomicrographs of the aortic wall on day 14 in the presence of SB203580, stained with EVG (upper row) or anti‐CD68 antibody (lower row). (h) Bar graph showing the number of CD68 + cells in the aortic wall on day 14 in the presence of SB203580 ( n = 5–6, p = 0.6623 [Mann–Whitney U test]). (i) Bar graphs showing the aorta diameter ( n = 5–6, p = 0.2468 [Mann–Whitney U test]), AAA incidence ( n = 5–6, p = 0.2506 [χ 2 test]), adventitia ( n = 5–6, p > 0.9999 [Mann–Whitney U test]), intramedia ( n = 5–6, p = 0.1710 [Mann–Whitney U test]), and elastin degradation score ( n = 5–6, p = 0.4917 [ t test], p = 0.6753 [Mann–Whitney U test]) on day 14 in the presence of SB203580.
    Figure Legend Snippet: The mechanism how interleukin‐38 suppresses macrophage activation. (a) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in the aortic wall on day 14. (b) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p = 0.0280 [ t test], p = 0.0411 [Mann–Whitney U test]) and p‐JNK ( n = 6, p = 0.6893 [ t test], p = 0.6991 [Mann–Whitney U test]). (c) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in macrophages after TNF‐α stimulation with or without IL‐38. (d) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p < 0.0001 [ANOVA], p < 0.0001 [post hoc, Control vs. TNF‐α], p = 0.0234 [post hoc, Control vs. TNF‐α + IL‐38], p = 0.0125 [post hoc, TNF‐α vs. TNF‐α + IL‐38]) and p‐JNK ( n = 6, p = 0.0283 [ANOVA], p = 0.0264 [post hoc, Control vs. TNF‐α]). (e) Representative western blot analysis of iNOS, MMP‐9, MMP‐2, and GAPDH in macrophages after TNF‐α stimulation with or without IL‐38 in the presence of SB203580. (f) Bar graphs showing the relative densitometry of iNOS ( n = 3, p = 0.2000 [Mann–Whitney U test]), MMP‐9 ( n = 3, p = 0.7000 [Mann–Whitney U test]), and MMP‐2 ( n = 3, p = 0.7000 [Mann–Whitney U test]). (g) Representative photomicrographs of the aortic wall on day 14 in the presence of SB203580, stained with EVG (upper row) or anti‐CD68 antibody (lower row). (h) Bar graph showing the number of CD68 + cells in the aortic wall on day 14 in the presence of SB203580 ( n = 5–6, p = 0.6623 [Mann–Whitney U test]). (i) Bar graphs showing the aorta diameter ( n = 5–6, p = 0.2468 [Mann–Whitney U test]), AAA incidence ( n = 5–6, p = 0.2506 [χ 2 test]), adventitia ( n = 5–6, p > 0.9999 [Mann–Whitney U test]), intramedia ( n = 5–6, p = 0.1710 [Mann–Whitney U test]), and elastin degradation score ( n = 5–6, p = 0.4917 [ t test], p = 0.6753 [Mann–Whitney U test]) on day 14 in the presence of SB203580.

    Techniques Used: Activation Assay, Western Blot, MANN-WHITNEY, Staining

    anti phospho p38 mapk  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38 mapk
    (A) Protein expression of different human PKC isoforms in the membrane and cytosol fractions of different HEK293 transfected cell lines (with or without treatment of ANGII). GC-A and NaK-ATPase serve as quality control for the separation of membrane and cytosol fractions. GAPDH serves as loading control. (B) Protein expression of phosphorylated PKC substrates, phosphorylated <t>p38</t> <t>MAPK,</t> and <t>p38</t> <t>MAPK</t> in HEK293/GC-A + /AT 1 + cells treated with different PKC modulators. GAPDH serves as loading control. PMA, phorbol 12-myristate 13-acetate. (C) In vitro cGMP generation in HEK293/GC-A + /AT 1 + in response to ANP (10 −8 M) with or without ANGII (10 −8 M) and valsartan (1 μM) or Go6983 (5 μM). Values of cGMP were normalized to vehicle group (blue) under ANP treatment. * indicates P <0.05, one-way ANOVA with Dunnett multiple comparisons test. (D) Hypothetical mechanism underlying the crosstalk between ANGII and GC-A derived from our in vitro studies.
    Anti Phospho P38 Mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "EVIDENCE FOR ANGIOTENSIN II AS A NATURALLY EXISTING SUPPRESSOR FOR THE NATRIURETIC PEPTIDE SYSTEM"

    Article Title: EVIDENCE FOR ANGIOTENSIN II AS A NATURALLY EXISTING SUPPRESSOR FOR THE NATRIURETIC PEPTIDE SYSTEM

    Journal: bioRxiv

    doi: 10.1101/2023.01.26.525806

    (A) Protein expression of different human PKC isoforms in the membrane and cytosol fractions of different HEK293 transfected cell lines (with or without treatment of ANGII). GC-A and NaK-ATPase serve as quality control for the separation of membrane and cytosol fractions. GAPDH serves as loading control. (B) Protein expression of phosphorylated PKC substrates, phosphorylated p38 MAPK, and p38 MAPK in HEK293/GC-A + /AT 1 + cells treated with different PKC modulators. GAPDH serves as loading control. PMA, phorbol 12-myristate 13-acetate. (C) In vitro cGMP generation in HEK293/GC-A + /AT 1 + in response to ANP (10 −8 M) with or without ANGII (10 −8 M) and valsartan (1 μM) or Go6983 (5 μM). Values of cGMP were normalized to vehicle group (blue) under ANP treatment. * indicates P <0.05, one-way ANOVA with Dunnett multiple comparisons test. (D) Hypothetical mechanism underlying the crosstalk between ANGII and GC-A derived from our in vitro studies.
    Figure Legend Snippet: (A) Protein expression of different human PKC isoforms in the membrane and cytosol fractions of different HEK293 transfected cell lines (with or without treatment of ANGII). GC-A and NaK-ATPase serve as quality control for the separation of membrane and cytosol fractions. GAPDH serves as loading control. (B) Protein expression of phosphorylated PKC substrates, phosphorylated p38 MAPK, and p38 MAPK in HEK293/GC-A + /AT 1 + cells treated with different PKC modulators. GAPDH serves as loading control. PMA, phorbol 12-myristate 13-acetate. (C) In vitro cGMP generation in HEK293/GC-A + /AT 1 + in response to ANP (10 −8 M) with or without ANGII (10 −8 M) and valsartan (1 μM) or Go6983 (5 μM). Values of cGMP were normalized to vehicle group (blue) under ANP treatment. * indicates P <0.05, one-way ANOVA with Dunnett multiple comparisons test. (D) Hypothetical mechanism underlying the crosstalk between ANGII and GC-A derived from our in vitro studies.

    Techniques Used: Expressing, Transfection, In Vitro, Derivative Assay

    phospho p38  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38
    Details of the antibodies used in this study.
    Phospho P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways"

    Article Title: Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways

    Journal: PLOS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0011062

    Details of the antibodies used in this study.
    Figure Legend Snippet: Details of the antibodies used in this study.

    Techniques Used:

    (A, B) The phosphorylation of AKT, p65, ERK and p38 in liver were analyzed by Western blot. (C, D) 3 × 10 5 WT and TLR2 -/- mouse BECs were cultivated with C . sinensis ESPs (50 μg/mL) for 120 min, the cells were lysed in cells lysates and the phosphorylation of AKT, p65, ERK and p38 were analyzed by Western blot. (E) The production levels of IL-6 in the supernatant of BECs pretreated with or without AKT inhibitor, p38 inhibitor and ERK inhibitor for 1 h, then co-cultured with ESPs for 18 h were measured by ELISA.
    Figure Legend Snippet: (A, B) The phosphorylation of AKT, p65, ERK and p38 in liver were analyzed by Western blot. (C, D) 3 × 10 5 WT and TLR2 -/- mouse BECs were cultivated with C . sinensis ESPs (50 μg/mL) for 120 min, the cells were lysed in cells lysates and the phosphorylation of AKT, p65, ERK and p38 were analyzed by Western blot. (E) The production levels of IL-6 in the supernatant of BECs pretreated with or without AKT inhibitor, p38 inhibitor and ERK inhibitor for 1 h, then co-cultured with ESPs for 18 h were measured by ELISA.

    Techniques Used: Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

    C . sinensis triggered AKT and p38 signal pathways dependent on TLR2 of BECs to promote the production of IL-6, which subsequently activated TGF-β1-Smad2/Smad3 pathways, myofibroblasts and up-regulated the expression of ECMs, finally resulting in biliary fibrosis in mice. In the figure, ‘ ↑ ’indicates positive regulation.
    Figure Legend Snippet: C . sinensis triggered AKT and p38 signal pathways dependent on TLR2 of BECs to promote the production of IL-6, which subsequently activated TGF-β1-Smad2/Smad3 pathways, myofibroblasts and up-regulated the expression of ECMs, finally resulting in biliary fibrosis in mice. In the figure, ‘ ↑ ’indicates positive regulation.

    Techniques Used: Expressing

    phospho p38  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38
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    rabbit anti phospho p38 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho p38 antibody
    Rabbit Anti Phospho P38 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho p38 mapk  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk
    A The indicated cell types were seeded on fibronectin-coated coverslips and stained with DAPI to visualize cell nuclei (blue) and with antibodies to alpha-tubulin (green) and gamma-tubulin (red). Shown are 3D confocal photographs generated using the Imaris software. Scale bars: 10 μm. B The X–Z axis stack projections of the photographs generated from laser-scanning confocal images taken in 0.3 μm steps were quantified using ZEN 2.3 software. The mitotic spindle angles of the indicated cell types were calculated. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. * P < 0.05, N = 10 (U87MG), N = 8 (clone 54), N = 11 (T98G ShC), N = 6 (T98G ShA2). C The cDNAs encoding A2InTm, A2ExTM or an empty expression vector (EV) were expressed in clone 54.3 knock-out cells (54.3). The phosphorylation levels of AKT were assayed in the indicated cell types by western blot analysis of cell lysates as described in methods, using an antibody directed against phosphorylated AKT (ser473). Loading was assessed using an antibody directed against total AKT. Shown is a representative western blot. The effect of A2InTm expression on the average phosphorylation levels of AKT was determined ( N = 8) (except for A2ExTm expressing cells, N = 1). Below is shown a histogram depicting the ratio between the intensity of phospho-AKT staining and total AKT. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. *** P < 0.001, D The phosphorylation levels of <t>p38</t> were assayed in the indicated cell types by western blot analysis of cell lysates as described in methods, using an antibody directed against phosphorylated p38 <t>(Thr180/Tyr182).</t> Loading was assessed using an antibody directed against total p38. Shown is a representative western blot. Below is shown a histogram depicting the average ratio between the intensity of the respective phospho-p38 bands and the total p38 bands. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. ** P < 0.01, N = 6.
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    1) Product Images from "Plexin-A2 enables the proliferation and the development of tumors from glioblastoma derived cells"

    Article Title: Plexin-A2 enables the proliferation and the development of tumors from glioblastoma derived cells

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-023-05554-0

    A The indicated cell types were seeded on fibronectin-coated coverslips and stained with DAPI to visualize cell nuclei (blue) and with antibodies to alpha-tubulin (green) and gamma-tubulin (red). Shown are 3D confocal photographs generated using the Imaris software. Scale bars: 10 μm. B The X–Z axis stack projections of the photographs generated from laser-scanning confocal images taken in 0.3 μm steps were quantified using ZEN 2.3 software. The mitotic spindle angles of the indicated cell types were calculated. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. * P < 0.05, N = 10 (U87MG), N = 8 (clone 54), N = 11 (T98G ShC), N = 6 (T98G ShA2). C The cDNAs encoding A2InTm, A2ExTM or an empty expression vector (EV) were expressed in clone 54.3 knock-out cells (54.3). The phosphorylation levels of AKT were assayed in the indicated cell types by western blot analysis of cell lysates as described in methods, using an antibody directed against phosphorylated AKT (ser473). Loading was assessed using an antibody directed against total AKT. Shown is a representative western blot. The effect of A2InTm expression on the average phosphorylation levels of AKT was determined ( N = 8) (except for A2ExTm expressing cells, N = 1). Below is shown a histogram depicting the ratio between the intensity of phospho-AKT staining and total AKT. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. *** P < 0.001, D The phosphorylation levels of p38 were assayed in the indicated cell types by western blot analysis of cell lysates as described in methods, using an antibody directed against phosphorylated p38 (Thr180/Tyr182). Loading was assessed using an antibody directed against total p38. Shown is a representative western blot. Below is shown a histogram depicting the average ratio between the intensity of the respective phospho-p38 bands and the total p38 bands. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. ** P < 0.01, N = 6.
    Figure Legend Snippet: A The indicated cell types were seeded on fibronectin-coated coverslips and stained with DAPI to visualize cell nuclei (blue) and with antibodies to alpha-tubulin (green) and gamma-tubulin (red). Shown are 3D confocal photographs generated using the Imaris software. Scale bars: 10 μm. B The X–Z axis stack projections of the photographs generated from laser-scanning confocal images taken in 0.3 μm steps were quantified using ZEN 2.3 software. The mitotic spindle angles of the indicated cell types were calculated. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. * P < 0.05, N = 10 (U87MG), N = 8 (clone 54), N = 11 (T98G ShC), N = 6 (T98G ShA2). C The cDNAs encoding A2InTm, A2ExTM or an empty expression vector (EV) were expressed in clone 54.3 knock-out cells (54.3). The phosphorylation levels of AKT were assayed in the indicated cell types by western blot analysis of cell lysates as described in methods, using an antibody directed against phosphorylated AKT (ser473). Loading was assessed using an antibody directed against total AKT. Shown is a representative western blot. The effect of A2InTm expression on the average phosphorylation levels of AKT was determined ( N = 8) (except for A2ExTm expressing cells, N = 1). Below is shown a histogram depicting the ratio between the intensity of phospho-AKT staining and total AKT. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. *** P < 0.001, D The phosphorylation levels of p38 were assayed in the indicated cell types by western blot analysis of cell lysates as described in methods, using an antibody directed against phosphorylated p38 (Thr180/Tyr182). Loading was assessed using an antibody directed against total p38. Shown is a representative western blot. Below is shown a histogram depicting the average ratio between the intensity of the respective phospho-p38 bands and the total p38 bands. Data are represented as mean ± SD. Statistical analysis was done using the one-tailed Mann-Whitney test. ** P < 0.01, N = 6.

    Techniques Used: Staining, Generated, Software, One-tailed Test, MANN-WHITNEY, Expressing, Plasmid Preparation, Knock-Out, Western Blot

    anti phospho p38  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38
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    Cell Signaling Technology Inc pathscan phospho p38 mapk thr180 tyr182 sandwich elisa
    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, <t>the</t> <t>p38-MAPK</t> inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases <t>in</t> <t>phospho-Thr180/Tyr182-p38</t> were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.
    Pathscan Phospho P38 Mapk Thr180 Tyr182 Sandwich Elisa, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho p38 mapk
    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, <t>the</t> <t>p38-MAPK</t> inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.
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    The mechanism how interleukin‐38 suppresses macrophage activation. (a) Representative western blot analysis of <t>p‐p38,</t> p38, p‐JNK, and JNK in the aortic wall on day 14. (b) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p = 0.0280 [ t test], p = 0.0411 [Mann–Whitney U test]) and p‐JNK ( n = 6, p = 0.6893 [ t test], p = 0.6991 [Mann–Whitney U test]). (c) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in macrophages after TNF‐α stimulation with or without IL‐38. (d) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p < 0.0001 [ANOVA], p < 0.0001 [post hoc, Control vs. TNF‐α], p = 0.0234 [post hoc, Control vs. TNF‐α + IL‐38], p = 0.0125 [post hoc, TNF‐α vs. TNF‐α + IL‐38]) and p‐JNK ( n = 6, p = 0.0283 [ANOVA], p = 0.0264 [post hoc, Control vs. TNF‐α]). (e) Representative western blot analysis of iNOS, MMP‐9, MMP‐2, and GAPDH in macrophages after TNF‐α stimulation with or without IL‐38 in the presence of SB203580. (f) Bar graphs showing the relative densitometry of iNOS ( n = 3, p = 0.2000 [Mann–Whitney U test]), MMP‐9 ( n = 3, p = 0.7000 [Mann–Whitney U test]), and MMP‐2 ( n = 3, p = 0.7000 [Mann–Whitney U test]). (g) Representative photomicrographs of the aortic wall on day 14 in the presence of SB203580, stained with EVG (upper row) or anti‐CD68 antibody (lower row). (h) Bar graph showing the number of CD68 + cells in the aortic wall on day 14 in the presence of SB203580 ( n = 5–6, p = 0.6623 [Mann–Whitney U test]). (i) Bar graphs showing the aorta diameter ( n = 5–6, p = 0.2468 [Mann–Whitney U test]), AAA incidence ( n = 5–6, p = 0.2506 [χ 2 test]), adventitia ( n = 5–6, p > 0.9999 [Mann–Whitney U test]), intramedia ( n = 5–6, p = 0.1710 [Mann–Whitney U test]), and elastin degradation score ( n = 5–6, p = 0.4917 [ t test], p = 0.6753 [Mann–Whitney U test]) on day 14 in the presence of SB203580.
    Anti Phospho P38 P P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho p38 mapk
    (A) Protein expression of different human PKC isoforms in the membrane and cytosol fractions of different HEK293 transfected cell lines (with or without treatment of ANGII). GC-A and NaK-ATPase serve as quality control for the separation of membrane and cytosol fractions. GAPDH serves as loading control. (B) Protein expression of phosphorylated PKC substrates, phosphorylated <t>p38</t> <t>MAPK,</t> and <t>p38</t> <t>MAPK</t> in HEK293/GC-A + /AT 1 + cells treated with different PKC modulators. GAPDH serves as loading control. PMA, phorbol 12-myristate 13-acetate. (C) In vitro cGMP generation in HEK293/GC-A + /AT 1 + in response to ANP (10 −8 M) with or without ANGII (10 −8 M) and valsartan (1 μM) or Go6983 (5 μM). Values of cGMP were normalized to vehicle group (blue) under ANP treatment. * indicates P <0.05, one-way ANOVA with Dunnett multiple comparisons test. (D) Hypothetical mechanism underlying the crosstalk between ANGII and GC-A derived from our in vitro studies.
    Anti Phospho P38 Mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Details of the antibodies used in this study.
    Phospho P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Details of the antibodies used in this study.
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    Cell Signaling Technology Inc anti phospho p38
    Details of the antibodies used in this study.
    Anti Phospho P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Expressing, Activity Assay, Activation Assay, Two Tailed Test, Standard Deviation

    a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Inhibition, Chromatin Immunoprecipitation, Agarose Gel Electrophoresis, Negative Control, Two Tailed Test, Standard Deviation

    A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Expressing, Two Tailed Test, Activity Assay

    Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Protein Binding, Activation Assay, Expressing, Binding Assay

    a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: a . The SPRBD- and SPRBD+IFNβ- induced increases in the mRNA expression of CHST15 and CHST11 were significantly reduced following exposure to SB20850, the p38-MAPK inhibitor. In contrast, NSC23766, a Rho/Rac-1 GTPase inhibitor, had no impact on their expression. b . SIS3, an inhibitor of phospho-Smad3, inhibited the SPRBD- and SPRBD+IFNβ- induced increases in expression of CHST15 and CHST11. c . Expression of ACE2 was inhibited by ACE2 specific siRNA following exposure to SPRBD alone or with IFNβ. d . Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD+IFNβ- induced increases in phospho-T180/T182-p38 MAPK were inhibited. e . The SPRBD- and SPRBD+IFNβ- induced increases in phospho-S423/S425 SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB20850, the p38 MAPK inhibitor. f . The SPRBD and SPRBD+IFNβ- induced increases in phospho-Thr180/Tyr182-p38 were unaffected by exposure to SIS3 and were inhibited by SB20850. g , h . Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFNβ and SPRBD. Both SIS3 and SB20850 inhibited promoter activation. [SIS3=specific inhibitor of SMAD3]. All p-values were determined by unpaired t-test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation.

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Expressing, Activity Assay, Activation Assay, Two Tailed Test, Standard Deviation

    a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: a . ARSB mRNA expression was unaffected by NSC23766, but following exposure to the p38-MAPK inhibitor SB20850, mRNA expression was restored to baseline control values. b . In contrast, SIS3 had no impact on the SPRBD- or SPRBD+IFNβ-induced decline in ARSB expression. c . ARSB promoter activation was reduced by SPRBD and by SPRBD+IFNβ. These declines were reversed by exposure to SB20850, but not by SIS3. d . Following treatment with SPRBD or SPRBD+IFNβ, C-terminal phospho-(Ser807/811)-Rb declined as shown by ELISA. These declines were reversed by SB203580, but not by SIS3. e . In contrast to the decline in C-terminal Rb phosphorylation, exposure to SPRBD+IFNβ increased N-terminus phospho-(S249)-Rb, as detected by Western blot. This increase was inhibited by SB20850, and total Rb was unchanged. f . Densitometry confirms the impression of Western blot and shows the ratio of phospho-S249-Rb to total Rb following SPRBD+IFNβ has increased to 3.89 times the baseline. g . Following exposure to SPRBD and SPRBD+IFNβ, E2F-DNA binding declined significantly, and SB20850 reversed the declines. h . %DNA input declined following SPRBD+IFNβ and increased following inhibition of p38-MAPK by SB20850. i . Chromatin immunoprecipitation (ChIP) shows decline in E2F1 binding to the ARSB promoter following exposure to SPRBD+IFNβ; decline was reversed by SB203580. Agarose gel demonstrates no effect of the IgG negative control on E2F1 binding to the ARSB promoter at baseline, IFNβ control, reduced binding following SPRBD+IFNβ, and reversal of the decline following treatment with SB20850. j . Measurements of optical density confirm the impression from the gel. P-values were determined by unpaired t-tests, two-tailed with unequal variance, with n of at least 3 independent experiments. Error bars show one standard deviation.

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Inhibition, Chromatin Immunoprecipitation, Agarose Gel Electrophoresis, Negative Control, Two Tailed Test, Standard Deviation

    A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: A . The antihistamine desloratadine reduced by 62% the SPRBD-induced increase in T180/T182 phospho-p38-MAPK in the AEC. B . Consistent with the decline in phospho-p38, desloratadine reduced the SPRBD-induced and SPRBD with IFNβ-induced increases in CHST15 (p=0.045, p=0.0041 with IFNβ) and CHST11 expression (p=0.008, p=0.0017 with IFNβ, unpaired t-tests, two-tailed, unequal variance, n=3). C, D . Consistent with the observed decline in ARSB, desloratadine partially reversed the SPRBD-induced decline in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFNβ; n=3) and expression (0.71 to 0.87 and 0.46 to 0.74 with IFNβ, fold-change compared to control; n=3). E . The polyether antibiotic monensin, which modifies dermatan sulfate biosynthesis and processing [ - ], reduced the SPRBD-induced increases in CHST15 mRNA (p=0.003, p=0.006 with IFNβ) and CHST11 (p=0.003, p=0.002 with IFNβ; fold-change compared to control, unpaired t-tests, two-tailed, unequal variance). F . Monensin had no significant impact on the SPRBD-induced decline in ARSB.

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Expressing, Two Tailed Test, Activity Assay

    Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]

    Journal: medRxiv

    Article Title: SARS-CoV-2 Spike Protein Receptor Binding-ACE2 Interaction Increases Carbohydrate Sulfotransferases and Reduces N-Acetylgalactosamine-4-Sulfatase through Phospho-p38-MAPK and RB-E2F1

    doi: 10.1101/2023.01.24.23284890

    Figure Lengend Snippet: Schematic of SARS-CoV-2 spike protein binding domain (SPRBD) peptide interaction and initiation of transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. Both p38 MAPK and phsopho-Smad3 lead to increased expression of CHST15 and CHST11. AngII interaction with AT1 is also expected to increase phospho-p38. Phospho-p38 leads to N-terminal phosphorylation and activation of Rb with enhanced binding of E2F1, which reduces ARSB promoter activation due to reduced binding of E2F1. [ACE2=angiotensin converting enzyme 2; AngII=angiotensin II; ARSB=arylsulfatase B; AT1R=angiotensin II receptor type 1; AT2R=angiotensin II receptor type 2; CHST=carbohydrate sulfotransferase; pRb=retinoblastoma protein; SMAD=Suppressor of Mothers against Decapentaplegic; SPRBD=SARS-CoV-2 spike protein receptor binding domain]

    Article Snippet: Phospho-p38 MAPK was measured in cell samples using a Pathscan® Phospho-p38 MAPK Thr180/Tyr182 Sandwich ELISA (Cell Signaling).

    Techniques: Protein Binding, Activation Assay, Expressing, Binding Assay

    The mechanism how interleukin‐38 suppresses macrophage activation. (a) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in the aortic wall on day 14. (b) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p = 0.0280 [ t test], p = 0.0411 [Mann–Whitney U test]) and p‐JNK ( n = 6, p = 0.6893 [ t test], p = 0.6991 [Mann–Whitney U test]). (c) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in macrophages after TNF‐α stimulation with or without IL‐38. (d) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p < 0.0001 [ANOVA], p < 0.0001 [post hoc, Control vs. TNF‐α], p = 0.0234 [post hoc, Control vs. TNF‐α + IL‐38], p = 0.0125 [post hoc, TNF‐α vs. TNF‐α + IL‐38]) and p‐JNK ( n = 6, p = 0.0283 [ANOVA], p = 0.0264 [post hoc, Control vs. TNF‐α]). (e) Representative western blot analysis of iNOS, MMP‐9, MMP‐2, and GAPDH in macrophages after TNF‐α stimulation with or without IL‐38 in the presence of SB203580. (f) Bar graphs showing the relative densitometry of iNOS ( n = 3, p = 0.2000 [Mann–Whitney U test]), MMP‐9 ( n = 3, p = 0.7000 [Mann–Whitney U test]), and MMP‐2 ( n = 3, p = 0.7000 [Mann–Whitney U test]). (g) Representative photomicrographs of the aortic wall on day 14 in the presence of SB203580, stained with EVG (upper row) or anti‐CD68 antibody (lower row). (h) Bar graph showing the number of CD68 + cells in the aortic wall on day 14 in the presence of SB203580 ( n = 5–6, p = 0.6623 [Mann–Whitney U test]). (i) Bar graphs showing the aorta diameter ( n = 5–6, p = 0.2468 [Mann–Whitney U test]), AAA incidence ( n = 5–6, p = 0.2506 [χ 2 test]), adventitia ( n = 5–6, p > 0.9999 [Mann–Whitney U test]), intramedia ( n = 5–6, p = 0.1710 [Mann–Whitney U test]), and elastin degradation score ( n = 5–6, p = 0.4917 [ t test], p = 0.6753 [Mann–Whitney U test]) on day 14 in the presence of SB203580.

    Journal: Physiological Reports

    Article Title: Interleukin‐38 suppresses abdominal aortic aneurysm formation in mice by regulating macrophages in an IL1RL2 ‐p38 pathway‐dependent manner

    doi: 10.14814/phy2.15581

    Figure Lengend Snippet: The mechanism how interleukin‐38 suppresses macrophage activation. (a) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in the aortic wall on day 14. (b) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p = 0.0280 [ t test], p = 0.0411 [Mann–Whitney U test]) and p‐JNK ( n = 6, p = 0.6893 [ t test], p = 0.6991 [Mann–Whitney U test]). (c) Representative western blot analysis of p‐p38, p38, p‐JNK, and JNK in macrophages after TNF‐α stimulation with or without IL‐38. (d) Bar graphs showing the relative densitometry of p‐p38 ( n = 6, p < 0.0001 [ANOVA], p < 0.0001 [post hoc, Control vs. TNF‐α], p = 0.0234 [post hoc, Control vs. TNF‐α + IL‐38], p = 0.0125 [post hoc, TNF‐α vs. TNF‐α + IL‐38]) and p‐JNK ( n = 6, p = 0.0283 [ANOVA], p = 0.0264 [post hoc, Control vs. TNF‐α]). (e) Representative western blot analysis of iNOS, MMP‐9, MMP‐2, and GAPDH in macrophages after TNF‐α stimulation with or without IL‐38 in the presence of SB203580. (f) Bar graphs showing the relative densitometry of iNOS ( n = 3, p = 0.2000 [Mann–Whitney U test]), MMP‐9 ( n = 3, p = 0.7000 [Mann–Whitney U test]), and MMP‐2 ( n = 3, p = 0.7000 [Mann–Whitney U test]). (g) Representative photomicrographs of the aortic wall on day 14 in the presence of SB203580, stained with EVG (upper row) or anti‐CD68 antibody (lower row). (h) Bar graph showing the number of CD68 + cells in the aortic wall on day 14 in the presence of SB203580 ( n = 5–6, p = 0.6623 [Mann–Whitney U test]). (i) Bar graphs showing the aorta diameter ( n = 5–6, p = 0.2468 [Mann–Whitney U test]), AAA incidence ( n = 5–6, p = 0.2506 [χ 2 test]), adventitia ( n = 5–6, p > 0.9999 [Mann–Whitney U test]), intramedia ( n = 5–6, p = 0.1710 [Mann–Whitney U test]), and elastin degradation score ( n = 5–6, p = 0.4917 [ t test], p = 0.6753 [Mann–Whitney U test]) on day 14 in the presence of SB203580.

    Article Snippet: Primary antibodies included anti‐TNF‐α (rabbit monoclonal, dilution 1:400, #11948; Cell Signaling Technology), anti‐iNOS (rabbit monoclonal, dilution 1:400, #13120; Cell Signaling Technology), anti‐ MMP‐9 (rabbit polyclonal, dilution 1:1000, #ab38898; abcam), anti‐ MMP‐2 (rabbit polyclonal, dilution 1:1000, #ab97779; abcam), anti‐IL1RL2 (rabbit polyclonal, dilution 1:1000, # PA5‐38013; Invitrogen), anti‐p38 (rabbit polyclonal, dilution 1:5000, #9212; Cell signaling technology), anti‐phospho‐p38 (p‐p38) (rabbit polyclonal, dilution 1:1000, #9211; Cell signaling technology), anti‐JNK (rabbit polyclonal, dilution 1:5000, #9252; Cell signaling technology) and anti‐phospho‐JNK (p‐JNK) (rabbit polyclonal, dilution 1:1000, #9251; Cell signaling technology).

    Techniques: Activation Assay, Western Blot, MANN-WHITNEY, Staining

    (A) Protein expression of different human PKC isoforms in the membrane and cytosol fractions of different HEK293 transfected cell lines (with or without treatment of ANGII). GC-A and NaK-ATPase serve as quality control for the separation of membrane and cytosol fractions. GAPDH serves as loading control. (B) Protein expression of phosphorylated PKC substrates, phosphorylated p38 MAPK, and p38 MAPK in HEK293/GC-A + /AT 1 + cells treated with different PKC modulators. GAPDH serves as loading control. PMA, phorbol 12-myristate 13-acetate. (C) In vitro cGMP generation in HEK293/GC-A + /AT 1 + in response to ANP (10 −8 M) with or without ANGII (10 −8 M) and valsartan (1 μM) or Go6983 (5 μM). Values of cGMP were normalized to vehicle group (blue) under ANP treatment. * indicates P <0.05, one-way ANOVA with Dunnett multiple comparisons test. (D) Hypothetical mechanism underlying the crosstalk between ANGII and GC-A derived from our in vitro studies.

    Journal: bioRxiv

    Article Title: EVIDENCE FOR ANGIOTENSIN II AS A NATURALLY EXISTING SUPPRESSOR FOR THE NATRIURETIC PEPTIDE SYSTEM

    doi: 10.1101/2023.01.26.525806

    Figure Lengend Snippet: (A) Protein expression of different human PKC isoforms in the membrane and cytosol fractions of different HEK293 transfected cell lines (with or without treatment of ANGII). GC-A and NaK-ATPase serve as quality control for the separation of membrane and cytosol fractions. GAPDH serves as loading control. (B) Protein expression of phosphorylated PKC substrates, phosphorylated p38 MAPK, and p38 MAPK in HEK293/GC-A + /AT 1 + cells treated with different PKC modulators. GAPDH serves as loading control. PMA, phorbol 12-myristate 13-acetate. (C) In vitro cGMP generation in HEK293/GC-A + /AT 1 + in response to ANP (10 −8 M) with or without ANGII (10 −8 M) and valsartan (1 μM) or Go6983 (5 μM). Values of cGMP were normalized to vehicle group (blue) under ANP treatment. * indicates P <0.05, one-way ANOVA with Dunnett multiple comparisons test. (D) Hypothetical mechanism underlying the crosstalk between ANGII and GC-A derived from our in vitro studies.

    Article Snippet: The following primary antibodies were used: anti-GCA (catalog#MAB48601, R&D System, Minneapolis, MN) at 1:1000 dilution, anti-GCB (catalog#55113-1-AP, Proteintech, Rosemont, IL) at 1:200 dilution, anti-GFP (catalog#TA150041, OriGene, Rockville, MD) at 1:1000 dilution, anti-PKCα (catalog#2056, Cell Signaling Technology, Danvers, MA) at 1:500 dilution, anti-PKCε (catalog#2683, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, anti-Phspho-(Ser) PKC Substrate (catalog#2261, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, anti-NaK-ATPase (catalog#EP1845Y, abcam, Waltham, MA) at 1:2000 dilution, anti-p38 MAPK (catalog#8690, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, anti-Phospho-p38 MAPK (catalog#4511, Cell Signaling Technology, Danvers, MA) at 1:1000 dilution, and anti-GAPDH (catalog#2118, Cell Signaling Technology, Danvers, MA) at 1:5000 dilution.

    Techniques: Expressing, Transfection, In Vitro, Derivative Assay

    Details of the antibodies used in this study.

    Journal: PLOS Neglected Tropical Diseases

    Article Title: Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways

    doi: 10.1371/journal.pntd.0011062

    Figure Lengend Snippet: Details of the antibodies used in this study.

    Article Snippet: Phospho-p38 , Rabbit monoclonal , Rabbit IgG , 1:1000 , Cell Signaling Technology.

    Techniques:

    (A, B) The phosphorylation of AKT, p65, ERK and p38 in liver were analyzed by Western blot. (C, D) 3 × 10 5 WT and TLR2 -/- mouse BECs were cultivated with C . sinensis ESPs (50 μg/mL) for 120 min, the cells were lysed in cells lysates and the phosphorylation of AKT, p65, ERK and p38 were analyzed by Western blot. (E) The production levels of IL-6 in the supernatant of BECs pretreated with or without AKT inhibitor, p38 inhibitor and ERK inhibitor for 1 h, then co-cultured with ESPs for 18 h were measured by ELISA.

    Journal: PLOS Neglected Tropical Diseases

    Article Title: Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways

    doi: 10.1371/journal.pntd.0011062

    Figure Lengend Snippet: (A, B) The phosphorylation of AKT, p65, ERK and p38 in liver were analyzed by Western blot. (C, D) 3 × 10 5 WT and TLR2 -/- mouse BECs were cultivated with C . sinensis ESPs (50 μg/mL) for 120 min, the cells were lysed in cells lysates and the phosphorylation of AKT, p65, ERK and p38 were analyzed by Western blot. (E) The production levels of IL-6 in the supernatant of BECs pretreated with or without AKT inhibitor, p38 inhibitor and ERK inhibitor for 1 h, then co-cultured with ESPs for 18 h were measured by ELISA.

    Article Snippet: Phospho-p38 , Rabbit monoclonal , Rabbit IgG , 1:1000 , Cell Signaling Technology.

    Techniques: Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

    C . sinensis triggered AKT and p38 signal pathways dependent on TLR2 of BECs to promote the production of IL-6, which subsequently activated TGF-β1-Smad2/Smad3 pathways, myofibroblasts and up-regulated the expression of ECMs, finally resulting in biliary fibrosis in mice. In the figure, ‘ ↑ ’indicates positive regulation.

    Journal: PLOS Neglected Tropical Diseases

    Article Title: Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways

    doi: 10.1371/journal.pntd.0011062

    Figure Lengend Snippet: C . sinensis triggered AKT and p38 signal pathways dependent on TLR2 of BECs to promote the production of IL-6, which subsequently activated TGF-β1-Smad2/Smad3 pathways, myofibroblasts and up-regulated the expression of ECMs, finally resulting in biliary fibrosis in mice. In the figure, ‘ ↑ ’indicates positive regulation.

    Article Snippet: Phospho-p38 , Rabbit monoclonal , Rabbit IgG , 1:1000 , Cell Signaling Technology.

    Techniques: Expressing