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Merck KGaA p38 inhibitor sb202190
<t>p38</t> inhibitor treatment reverses RNASE1 promoter deacetylation and CHD4 recruitment in inflamed human ECs. HUVEC were pretreated with 20 μM p38 inhibitor (inh.) SB202190 (+) or DMSO as solvent control (–) for 2 h, followed by 10 min TNF-α stimulation [10 ng/ml] (white bars) or left untreated as control (CTRL; black bars). Immunoprecipitation using specific antibodies against histone 4 acetylation (H4ac; left panels), histone 3 lysine 27 acetylation (H3K27ac; middle panels), chromodomain helicase DNA binding protein 4 (CHD4; right panels) or an unspecific IgG control was performed. (A) Region A , (B) Region B , (C) Region C of the RNASE1 promoter were pulled down by the respective antibodies and analyzed by qPCR using respective primers. Results were depicted as % input and the respective control sample with the specific antibody was set to 1. n = 3–4; mean ± SD; Two-way ANOVA was performed using Holm-Sidak post-test. *IgG vs. specific (spec.) antibody (AB): * p
P38 Inhibitor Sb202190, supplied by Merck KGaA, 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 "p38 and Casein Kinase 2 Mediate Ribonuclease 1 Repression in Inflamed Human Endothelial Cells via Promoter Remodeling Through Nucleosome Remodeling and Deacetylase Complex"

Article Title: p38 and Casein Kinase 2 Mediate Ribonuclease 1 Repression in Inflamed Human Endothelial Cells via Promoter Remodeling Through Nucleosome Remodeling and Deacetylase Complex

Journal: Frontiers in Cell and Developmental Biology

doi: 10.3389/fcell.2020.563604

p38 inhibitor treatment reverses RNASE1 promoter deacetylation and CHD4 recruitment in inflamed human ECs. HUVEC were pretreated with 20 μM p38 inhibitor (inh.) SB202190 (+) or DMSO as solvent control (–) for 2 h, followed by 10 min TNF-α stimulation [10 ng/ml] (white bars) or left untreated as control (CTRL; black bars). Immunoprecipitation using specific antibodies against histone 4 acetylation (H4ac; left panels), histone 3 lysine 27 acetylation (H3K27ac; middle panels), chromodomain helicase DNA binding protein 4 (CHD4; right panels) or an unspecific IgG control was performed. (A) Region A , (B) Region B , (C) Region C of the RNASE1 promoter were pulled down by the respective antibodies and analyzed by qPCR using respective primers. Results were depicted as % input and the respective control sample with the specific antibody was set to 1. n = 3–4; mean ± SD; Two-way ANOVA was performed using Holm-Sidak post-test. *IgG vs. specific (spec.) antibody (AB): * p
Figure Legend Snippet: p38 inhibitor treatment reverses RNASE1 promoter deacetylation and CHD4 recruitment in inflamed human ECs. HUVEC were pretreated with 20 μM p38 inhibitor (inh.) SB202190 (+) or DMSO as solvent control (–) for 2 h, followed by 10 min TNF-α stimulation [10 ng/ml] (white bars) or left untreated as control (CTRL; black bars). Immunoprecipitation using specific antibodies against histone 4 acetylation (H4ac; left panels), histone 3 lysine 27 acetylation (H3K27ac; middle panels), chromodomain helicase DNA binding protein 4 (CHD4; right panels) or an unspecific IgG control was performed. (A) Region A , (B) Region B , (C) Region C of the RNASE1 promoter were pulled down by the respective antibodies and analyzed by qPCR using respective primers. Results were depicted as % input and the respective control sample with the specific antibody was set to 1. n = 3–4; mean ± SD; Two-way ANOVA was performed using Holm-Sidak post-test. *IgG vs. specific (spec.) antibody (AB): * p

Techniques Used: Immunoprecipitation, Binding Assay, Real-time Polymerase Chain Reaction

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    Merck KGaA sb202190
    Involvement of the p38 and JNK MAPK pathway in 15d-PGJ 2 -induced cell death. Effects of a p38 MAPK inhibitor, <t>SB202190</t> (A), or JNK inhibitor, SP600125 (B, C), on 15d-PGJ 2 -induced cell death and the effects of 15d-PGJ 2 on phosphorylation of JNK (D). Cells were precultured for 24 h at 5 × 10 3 /well in 96-well plates and exposed to 15d-PGJ 2 at approximately the IC 50 in the presence or absence of SB202190 (3 μM) or SP600125 (0.1, 0.3, 1 μM) for 24 h. Cell viability was assessed by fluorescent assay, and data represent the mean ± S.D. from 4 independent preparations. Statistical significance was assessed by t-test or Dunnett's test. To detect proteins, cells were precultured for 24 h in 100-mm dishes. Cells were then treated with 15d-PGJ 2 at 3 μM for 0 and 8 h. Protein (15 μg) was analyzed by Western blotting for the expression of phospho-JNK. Relative protein levels were quantified using ImageJ, and each phospho-JNK signal was normalized to the β-actin signal. The representative bands and the results of densitometric analysis from three independent preparations were described, and data represent the mean ± S.D. from 4 independent preparations.
    Sb202190, supplied by Merck KGaA, 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/sb202190/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sb202190 - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

    86
    Merck KGaA p38 inhibitor sb202190
    <t>p38</t> inhibitor treatment reverses RNASE1 promoter deacetylation and CHD4 recruitment in inflamed human ECs. HUVEC were pretreated with 20 μM p38 inhibitor (inh.) SB202190 (+) or DMSO as solvent control (–) for 2 h, followed by 10 min TNF-α stimulation [10 ng/ml] (white bars) or left untreated as control (CTRL; black bars). Immunoprecipitation using specific antibodies against histone 4 acetylation (H4ac; left panels), histone 3 lysine 27 acetylation (H3K27ac; middle panels), chromodomain helicase DNA binding protein 4 (CHD4; right panels) or an unspecific IgG control was performed. (A) Region A , (B) Region B , (C) Region C of the RNASE1 promoter were pulled down by the respective antibodies and analyzed by qPCR using respective primers. Results were depicted as % input and the respective control sample with the specific antibody was set to 1. n = 3–4; mean ± SD; Two-way ANOVA was performed using Holm-Sidak post-test. *IgG vs. specific (spec.) antibody (AB): * p
    P38 Inhibitor Sb202190, supplied by Merck KGaA, 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/p38 inhibitor sb202190/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p38 inhibitor sb202190 - by Bioz Stars, 2021-04
    86/100 stars
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    86
    Merck KGaA p38 inhibitor
    Apoptosis caused by overexpression of DUSP6 and DUSP8. (A and B) LCLs were overexpressed with wild-type DUSP6 (DUSP6 WT), wild-type DUSP8 (DUSP8 WT) or their catalytic mutants (DUSP6 CS and DUSP8 CS). Selected cells underwent PI staining, and cell cycle patterns were analyzed with flow cytometry. Cells were selected with G418, and their lysates were analyzed by Western blotting. (C) Protein expression of DUSP6, phosphorylated ERK (p-ERK), total ERK (t-ERK), caspase-3, cleaved caspase-3, PARP-1, and cleaved PARP-1 was detected by immunoblotting. β-Actin served as an internal control. (D) Protein expression of DUSP8, phosphorylated <t>p38</t> (p-p38), total p38 (t-p38), phosphorylated JNK (p-JNK), and total JNK (t-JNK) was detected. β-Actin served as an internal control. (E) Protein expression of DUSP8, phosphorylated p38 (p-p38), total p38 (t-p38), and PUMA was detected by Western blotting. β-Actin served as the internal control. DUSP6 and DUSP8 were transduced into LCLs through lentiviral infection, and the cells were selected with G418 for 5 days. (A and B) Selected cells underwent PI staining, and cell cycle patterns were analyzed with flow cytometry. (C to E) LCLs were overexpressed with wild-type DUSP6 (DUSP6 WT) or wild-type DUSP8 (DUSP8 WT) or with their catalytic mutants (DUSP6 CS and DUSP8 CS). Cells were selected with G418, and their lysates were analyzed by Western blotting. (C) Protein expression of DUSP6, phosphorylated ERK (p-ERK), total ERK (t-ERK), caspase-3, cleaved caspase-3, PARP-1, and cleaved PARP-1 was detected by immunoblotting. β-Actin served as an internal control. (D) Protein expression of WT DUSP8, phosphorylated ERK (p-ERK), total ERK (t-ERK), phosphorylated JNK (p-JNK), total JNK (t-JNK), phosphorylated p38 (p-p38), and total p38 (t-p38) was detected. β-Actin served as an internal control. (E) Protein expression of wild-type DUSP8, DUSP8 CS, phosphorylated p38 (p-p38), total p38 (t-p38), and PUMA was detected by Western blotting. β-Actin served as the internal control.
    P38 Inhibitor, supplied by Merck KGaA, 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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    86
    Merck KGaA p38 mapk inhibitor sb202190
    Targeting of Dsg3 leads to <t>p38</t> <t>MAPK-dependent</t> loss of cell cohesion. A , targeting of Dsg3 function by either incubation with AK23 or siRNA-mediated depletion of Dsg3 protein levels in HaCaT cells resulted in increased cell monolayer fragmentation in dispase-based
    P38 Mapk Inhibitor Sb202190, supplied by Merck KGaA, 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/p38 mapk inhibitor sb202190/product/Merck KGaA
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    Involvement of the p38 and JNK MAPK pathway in 15d-PGJ 2 -induced cell death. Effects of a p38 MAPK inhibitor, SB202190 (A), or JNK inhibitor, SP600125 (B, C), on 15d-PGJ 2 -induced cell death and the effects of 15d-PGJ 2 on phosphorylation of JNK (D). Cells were precultured for 24 h at 5 × 10 3 /well in 96-well plates and exposed to 15d-PGJ 2 at approximately the IC 50 in the presence or absence of SB202190 (3 μM) or SP600125 (0.1, 0.3, 1 μM) for 24 h. Cell viability was assessed by fluorescent assay, and data represent the mean ± S.D. from 4 independent preparations. Statistical significance was assessed by t-test or Dunnett's test. To detect proteins, cells were precultured for 24 h in 100-mm dishes. Cells were then treated with 15d-PGJ 2 at 3 μM for 0 and 8 h. Protein (15 μg) was analyzed by Western blotting for the expression of phospho-JNK. Relative protein levels were quantified using ImageJ, and each phospho-JNK signal was normalized to the β-actin signal. The representative bands and the results of densitometric analysis from three independent preparations were described, and data represent the mean ± S.D. from 4 independent preparations.

    Journal: International Journal of Medical Sciences

    Article Title: Cytotoxicity of 15-Deoxy-?12,14-prostaglandin J2 through PPAR?-independent Pathway and the Involvement of the JNK and Akt Pathway in Renal Cell Carcinoma

    doi: 10.7150/ijms.4455

    Figure Lengend Snippet: Involvement of the p38 and JNK MAPK pathway in 15d-PGJ 2 -induced cell death. Effects of a p38 MAPK inhibitor, SB202190 (A), or JNK inhibitor, SP600125 (B, C), on 15d-PGJ 2 -induced cell death and the effects of 15d-PGJ 2 on phosphorylation of JNK (D). Cells were precultured for 24 h at 5 × 10 3 /well in 96-well plates and exposed to 15d-PGJ 2 at approximately the IC 50 in the presence or absence of SB202190 (3 μM) or SP600125 (0.1, 0.3, 1 μM) for 24 h. Cell viability was assessed by fluorescent assay, and data represent the mean ± S.D. from 4 independent preparations. Statistical significance was assessed by t-test or Dunnett's test. To detect proteins, cells were precultured for 24 h in 100-mm dishes. Cells were then treated with 15d-PGJ 2 at 3 μM for 0 and 8 h. Protein (15 μg) was analyzed by Western blotting for the expression of phospho-JNK. Relative protein levels were quantified using ImageJ, and each phospho-JNK signal was normalized to the β-actin signal. The representative bands and the results of densitometric analysis from three independent preparations were described, and data represent the mean ± S.D. from 4 independent preparations.

    Article Snippet: SB202190 (p38 inhibitor), SP600125 (c-Jun N-terminal kinase (JNK) inhibitor), and Akt inhibitor IV were obtained from Merck (Darmstadt, Germany).

    Techniques: Fluorescence, Western Blot, Expressing

    p38 inhibitor treatment reverses RNASE1 promoter deacetylation and CHD4 recruitment in inflamed human ECs. HUVEC were pretreated with 20 μM p38 inhibitor (inh.) SB202190 (+) or DMSO as solvent control (–) for 2 h, followed by 10 min TNF-α stimulation [10 ng/ml] (white bars) or left untreated as control (CTRL; black bars). Immunoprecipitation using specific antibodies against histone 4 acetylation (H4ac; left panels), histone 3 lysine 27 acetylation (H3K27ac; middle panels), chromodomain helicase DNA binding protein 4 (CHD4; right panels) or an unspecific IgG control was performed. (A) Region A , (B) Region B , (C) Region C of the RNASE1 promoter were pulled down by the respective antibodies and analyzed by qPCR using respective primers. Results were depicted as % input and the respective control sample with the specific antibody was set to 1. n = 3–4; mean ± SD; Two-way ANOVA was performed using Holm-Sidak post-test. *IgG vs. specific (spec.) antibody (AB): * p

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: p38 and Casein Kinase 2 Mediate Ribonuclease 1 Repression in Inflamed Human Endothelial Cells via Promoter Remodeling Through Nucleosome Remodeling and Deacetylase Complex

    doi: 10.3389/fcell.2020.563604

    Figure Lengend Snippet: p38 inhibitor treatment reverses RNASE1 promoter deacetylation and CHD4 recruitment in inflamed human ECs. HUVEC were pretreated with 20 μM p38 inhibitor (inh.) SB202190 (+) or DMSO as solvent control (–) for 2 h, followed by 10 min TNF-α stimulation [10 ng/ml] (white bars) or left untreated as control (CTRL; black bars). Immunoprecipitation using specific antibodies against histone 4 acetylation (H4ac; left panels), histone 3 lysine 27 acetylation (H3K27ac; middle panels), chromodomain helicase DNA binding protein 4 (CHD4; right panels) or an unspecific IgG control was performed. (A) Region A , (B) Region B , (C) Region C of the RNASE1 promoter were pulled down by the respective antibodies and analyzed by qPCR using respective primers. Results were depicted as % input and the respective control sample with the specific antibody was set to 1. n = 3–4; mean ± SD; Two-way ANOVA was performed using Holm-Sidak post-test. *IgG vs. specific (spec.) antibody (AB): * p

    Article Snippet: For inhibitor assays, HUVEC were pretreated for 1 h with the NF-κB inhibitor BAY11-7082 [1 μM, 5 μM], the JNK inhibitor JNK inhibitor II [10 μM, 30 μM], the p38 inhibitor SB202190 [10 μM, 20 μM] (Merck KGaA, Sigma Aldrich, Darmstadt, HE, Germany) prior to indicated stimulation for 24 h. Dimethyl sulfoxide (DMSO) (Carl Roth GmbH & Co., KG, Karlsruhe, BW, Germany) was used as solvent control.

    Techniques: Immunoprecipitation, Binding Assay, Real-time Polymerase Chain Reaction

    Apoptosis caused by overexpression of DUSP6 and DUSP8. (A and B) LCLs were overexpressed with wild-type DUSP6 (DUSP6 WT), wild-type DUSP8 (DUSP8 WT) or their catalytic mutants (DUSP6 CS and DUSP8 CS). Selected cells underwent PI staining, and cell cycle patterns were analyzed with flow cytometry. Cells were selected with G418, and their lysates were analyzed by Western blotting. (C) Protein expression of DUSP6, phosphorylated ERK (p-ERK), total ERK (t-ERK), caspase-3, cleaved caspase-3, PARP-1, and cleaved PARP-1 was detected by immunoblotting. β-Actin served as an internal control. (D) Protein expression of DUSP8, phosphorylated p38 (p-p38), total p38 (t-p38), phosphorylated JNK (p-JNK), and total JNK (t-JNK) was detected. β-Actin served as an internal control. (E) Protein expression of DUSP8, phosphorylated p38 (p-p38), total p38 (t-p38), and PUMA was detected by Western blotting. β-Actin served as the internal control. DUSP6 and DUSP8 were transduced into LCLs through lentiviral infection, and the cells were selected with G418 for 5 days. (A and B) Selected cells underwent PI staining, and cell cycle patterns were analyzed with flow cytometry. (C to E) LCLs were overexpressed with wild-type DUSP6 (DUSP6 WT) or wild-type DUSP8 (DUSP8 WT) or with their catalytic mutants (DUSP6 CS and DUSP8 CS). Cells were selected with G418, and their lysates were analyzed by Western blotting. (C) Protein expression of DUSP6, phosphorylated ERK (p-ERK), total ERK (t-ERK), caspase-3, cleaved caspase-3, PARP-1, and cleaved PARP-1 was detected by immunoblotting. β-Actin served as an internal control. (D) Protein expression of WT DUSP8, phosphorylated ERK (p-ERK), total ERK (t-ERK), phosphorylated JNK (p-JNK), total JNK (t-JNK), phosphorylated p38 (p-p38), and total p38 (t-p38) was detected. β-Actin served as an internal control. (E) Protein expression of wild-type DUSP8, DUSP8 CS, phosphorylated p38 (p-p38), total p38 (t-p38), and PUMA was detected by Western blotting. β-Actin served as the internal control.

    Journal: Journal of Virology

    Article Title: Dysregulation of Dual-Specificity Phosphatases by Epstein-Barr Virus LMP1 and Its Impact on Lymphoblastoid Cell Line Survival

    doi: 10.1128/JVI.01837-19

    Figure Lengend Snippet: Apoptosis caused by overexpression of DUSP6 and DUSP8. (A and B) LCLs were overexpressed with wild-type DUSP6 (DUSP6 WT), wild-type DUSP8 (DUSP8 WT) or their catalytic mutants (DUSP6 CS and DUSP8 CS). Selected cells underwent PI staining, and cell cycle patterns were analyzed with flow cytometry. Cells were selected with G418, and their lysates were analyzed by Western blotting. (C) Protein expression of DUSP6, phosphorylated ERK (p-ERK), total ERK (t-ERK), caspase-3, cleaved caspase-3, PARP-1, and cleaved PARP-1 was detected by immunoblotting. β-Actin served as an internal control. (D) Protein expression of DUSP8, phosphorylated p38 (p-p38), total p38 (t-p38), phosphorylated JNK (p-JNK), and total JNK (t-JNK) was detected. β-Actin served as an internal control. (E) Protein expression of DUSP8, phosphorylated p38 (p-p38), total p38 (t-p38), and PUMA was detected by Western blotting. β-Actin served as the internal control. DUSP6 and DUSP8 were transduced into LCLs through lentiviral infection, and the cells were selected with G418 for 5 days. (A and B) Selected cells underwent PI staining, and cell cycle patterns were analyzed with flow cytometry. (C to E) LCLs were overexpressed with wild-type DUSP6 (DUSP6 WT) or wild-type DUSP8 (DUSP8 WT) or with their catalytic mutants (DUSP6 CS and DUSP8 CS). Cells were selected with G418, and their lysates were analyzed by Western blotting. (C) Protein expression of DUSP6, phosphorylated ERK (p-ERK), total ERK (t-ERK), caspase-3, cleaved caspase-3, PARP-1, and cleaved PARP-1 was detected by immunoblotting. β-Actin served as an internal control. (D) Protein expression of WT DUSP8, phosphorylated ERK (p-ERK), total ERK (t-ERK), phosphorylated JNK (p-JNK), total JNK (t-JNK), phosphorylated p38 (p-p38), and total p38 (t-p38) was detected. β-Actin served as an internal control. (E) Protein expression of wild-type DUSP8, DUSP8 CS, phosphorylated p38 (p-p38), total p38 (t-p38), and PUMA was detected by Western blotting. β-Actin served as the internal control.

    Article Snippet: MEK inhibitor (U0126), phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002), IκB-α phosphorylation inhibitor (BAY11-7082), JNK inhibitor (SP600125), and p38 inhibitor (SB202190) were purchased from Merck Millipore (Billerica, MA, USA).

    Techniques: Over Expression, Staining, Flow Cytometry, Western Blot, Expressing, Infection

    Signaling pathways involved in LMP1-mediated DUSP6 and DUSP8 suppression. (A) LCLs were treated with dimethyl sulfoxide (DMSO), 20 μM U0126 (ERK inhibitor), 20 μM SB212090 (p38 inhibitor), 25 μM SP600125 (JNK inhibitor), 20 μM LY294002 (Akt inhibitor), or 10 μM Bay11-7082 (NF-κB inhibitor) for 2 days. Detection of DUSP6, DUSP8, phosphorylated ERK (p-ERK), phosphorylated p38 (p-p38), phosphorylated JNK (p-JNK), phosphorylated Akt (p-Akt), phosphorylated IκB (p-IκB), total ERK (t-ERK), total p38 (t-p38), total JNK (t-JNK), total Akt (t-Akt), total IκB (t-IκB), and LMP1 was carried out by Western blotting. β-Actin served as an internal control. The fold relative expression data were calculated by comparing the levels of β-actin-normalized DUSP expression of the inhibitor-treated LCLs with those of the untreated LCLs. (B) BJAB cells were transduced with vector control (pSIN) or LMP1 via lentiviral infection and treated with DMSO, 20 μM U0126, or 20 μM SB212090 for 2 days. Detection of DUSP6, DUSP8, p-ERK, p-p38, t-ERK, t-p38, and LMP1 was carried out by Western blotting. The related level of expression of DUSP in vector control-transfected, untreated cells was set as a value of 1 to calculate the fold relative expression levels of DUSP6 and DUSP8.

    Journal: Journal of Virology

    Article Title: Dysregulation of Dual-Specificity Phosphatases by Epstein-Barr Virus LMP1 and Its Impact on Lymphoblastoid Cell Line Survival

    doi: 10.1128/JVI.01837-19

    Figure Lengend Snippet: Signaling pathways involved in LMP1-mediated DUSP6 and DUSP8 suppression. (A) LCLs were treated with dimethyl sulfoxide (DMSO), 20 μM U0126 (ERK inhibitor), 20 μM SB212090 (p38 inhibitor), 25 μM SP600125 (JNK inhibitor), 20 μM LY294002 (Akt inhibitor), or 10 μM Bay11-7082 (NF-κB inhibitor) for 2 days. Detection of DUSP6, DUSP8, phosphorylated ERK (p-ERK), phosphorylated p38 (p-p38), phosphorylated JNK (p-JNK), phosphorylated Akt (p-Akt), phosphorylated IκB (p-IκB), total ERK (t-ERK), total p38 (t-p38), total JNK (t-JNK), total Akt (t-Akt), total IκB (t-IκB), and LMP1 was carried out by Western blotting. β-Actin served as an internal control. The fold relative expression data were calculated by comparing the levels of β-actin-normalized DUSP expression of the inhibitor-treated LCLs with those of the untreated LCLs. (B) BJAB cells were transduced with vector control (pSIN) or LMP1 via lentiviral infection and treated with DMSO, 20 μM U0126, or 20 μM SB212090 for 2 days. Detection of DUSP6, DUSP8, p-ERK, p-p38, t-ERK, t-p38, and LMP1 was carried out by Western blotting. The related level of expression of DUSP in vector control-transfected, untreated cells was set as a value of 1 to calculate the fold relative expression levels of DUSP6 and DUSP8.

    Article Snippet: MEK inhibitor (U0126), phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002), IκB-α phosphorylation inhibitor (BAY11-7082), JNK inhibitor (SP600125), and p38 inhibitor (SB202190) were purchased from Merck Millipore (Billerica, MA, USA).

    Techniques: Western Blot, Expressing, Transduction, Plasmid Preparation, Infection, Transfection

    Kinetic expression of MAPK activation and DUSPs during EBV immortalization. Human CD19 + B lymphocytes were purified from the buffy coat of a healthy blood donor. Cells were seeded in a 12-well plate at a density of 1 × 10 6 cells per well and infected with EBV. At 28 days postinfection, LCLs were established. RNA and proteins were harvested at the time points indicated. (A) Protein expression of MAPK activation, including phosphorylation of ERK (p-ERK), p38 (p-p38), and JNK (p-JNK), during EBV immortalization was detected by Western blotting. Total expression of MAPK (t-ERK, t-p38, and t-JNK) also is shown. β-Actin was the internal control. (B to E) Cells were also harvested for RT-Q-PCR for expression of DUSPs. The relative expression levels of DUSPs were compared to those of uninfected B lymphocytes after normalization with GAPDH (glyceraldehyde-3-phosphate dehydrogenase) expression. The DUSPs analyzed included (B) typical type I DUSPs (DUSP1, DUSP2, DUSP4, and DUSP5), (C) typical type II DUSPs (DUSP6, DUSP7, and DUSP9), (D) typical type III DUSPs (DUSP8, DUSP10, and DUSP16), and (E) atypical DUSPs (DUSP14, DUSP19, and DUSP22).

    Journal: Journal of Virology

    Article Title: Dysregulation of Dual-Specificity Phosphatases by Epstein-Barr Virus LMP1 and Its Impact on Lymphoblastoid Cell Line Survival

    doi: 10.1128/JVI.01837-19

    Figure Lengend Snippet: Kinetic expression of MAPK activation and DUSPs during EBV immortalization. Human CD19 + B lymphocytes were purified from the buffy coat of a healthy blood donor. Cells were seeded in a 12-well plate at a density of 1 × 10 6 cells per well and infected with EBV. At 28 days postinfection, LCLs were established. RNA and proteins were harvested at the time points indicated. (A) Protein expression of MAPK activation, including phosphorylation of ERK (p-ERK), p38 (p-p38), and JNK (p-JNK), during EBV immortalization was detected by Western blotting. Total expression of MAPK (t-ERK, t-p38, and t-JNK) also is shown. β-Actin was the internal control. (B to E) Cells were also harvested for RT-Q-PCR for expression of DUSPs. The relative expression levels of DUSPs were compared to those of uninfected B lymphocytes after normalization with GAPDH (glyceraldehyde-3-phosphate dehydrogenase) expression. The DUSPs analyzed included (B) typical type I DUSPs (DUSP1, DUSP2, DUSP4, and DUSP5), (C) typical type II DUSPs (DUSP6, DUSP7, and DUSP9), (D) typical type III DUSPs (DUSP8, DUSP10, and DUSP16), and (E) atypical DUSPs (DUSP14, DUSP19, and DUSP22).

    Article Snippet: MEK inhibitor (U0126), phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002), IκB-α phosphorylation inhibitor (BAY11-7082), JNK inhibitor (SP600125), and p38 inhibitor (SB202190) were purchased from Merck Millipore (Billerica, MA, USA).

    Techniques: Expressing, Activation Assay, Purification, Infection, Western Blot, Polymerase Chain Reaction

    Targeting of Dsg3 leads to p38 MAPK-dependent loss of cell cohesion. A , targeting of Dsg3 function by either incubation with AK23 or siRNA-mediated depletion of Dsg3 protein levels in HaCaT cells resulted in increased cell monolayer fragmentation in dispase-based

    Journal: The Journal of Biological Chemistry

    Article Title: Desmoglein 2 Compensates for Desmoglein 3 but Does Not Control Cell Adhesion via Regulation of p38 Mitogen-activated Protein Kinase in Keratinocytes

    doi: 10.1074/jbc.M113.489336

    Figure Lengend Snippet: Targeting of Dsg3 leads to p38 MAPK-dependent loss of cell cohesion. A , targeting of Dsg3 function by either incubation with AK23 or siRNA-mediated depletion of Dsg3 protein levels in HaCaT cells resulted in increased cell monolayer fragmentation in dispase-based

    Article Snippet: Strikingly, 1 h of preincubation with the p38 MAPK inhibitor SB202190 (3.4 ± 1.2) or 24-h incubation of Dsg3-depleted cells with SB202190 significantly reduced the number of fragments (5.6 ± 1.5).

    Techniques: Incubation

    p38 MAPK is activated following Dsg3 depletion and forms a complex with Dsg3. A , HaCaT cells were transfected with siRNA targeting either Dsg2 or Dsg3, and successful knockdown was proven by reduced protein levels in a Western blot analysis against non-targeting

    Journal: The Journal of Biological Chemistry

    Article Title: Desmoglein 2 Compensates for Desmoglein 3 but Does Not Control Cell Adhesion via Regulation of p38 Mitogen-activated Protein Kinase in Keratinocytes

    doi: 10.1074/jbc.M113.489336

    Figure Lengend Snippet: p38 MAPK is activated following Dsg3 depletion and forms a complex with Dsg3. A , HaCaT cells were transfected with siRNA targeting either Dsg2 or Dsg3, and successful knockdown was proven by reduced protein levels in a Western blot analysis against non-targeting

    Article Snippet: Strikingly, 1 h of preincubation with the p38 MAPK inhibitor SB202190 (3.4 ± 1.2) or 24-h incubation of Dsg3-depleted cells with SB202190 significantly reduced the number of fragments (5.6 ± 1.5).

    Techniques: Transfection, Western Blot

    Dsg3 forms a complex with p-p38 MAPK in both the Triton X-100-soluble and Triton X-100-insoluble pool. Western blot analysis following Triton X-100-mediated cell fractionation in HaCaT cells revealed that Dsg3 was present in the soluble ( S ) and insoluble

    Journal: The Journal of Biological Chemistry

    Article Title: Desmoglein 2 Compensates for Desmoglein 3 but Does Not Control Cell Adhesion via Regulation of p38 Mitogen-activated Protein Kinase in Keratinocytes

    doi: 10.1074/jbc.M113.489336

    Figure Lengend Snippet: Dsg3 forms a complex with p-p38 MAPK in both the Triton X-100-soluble and Triton X-100-insoluble pool. Western blot analysis following Triton X-100-mediated cell fractionation in HaCaT cells revealed that Dsg3 was present in the soluble ( S ) and insoluble

    Article Snippet: Strikingly, 1 h of preincubation with the p38 MAPK inhibitor SB202190 (3.4 ± 1.2) or 24-h incubation of Dsg3-depleted cells with SB202190 significantly reduced the number of fragments (5.6 ± 1.5).

    Techniques: Western Blot, Cell Fractionation

    Mechanisms of Dsg2- and Dsg3-mediated cell-cell adhesion in keratinocytes. Targeting of Dsg3 binding either by pemphigus antibodies or siRNA leads to p38 MAPK activation and following retraction of the keratin filament network, which both negatively affect

    Journal: The Journal of Biological Chemistry

    Article Title: Desmoglein 2 Compensates for Desmoglein 3 but Does Not Control Cell Adhesion via Regulation of p38 Mitogen-activated Protein Kinase in Keratinocytes

    doi: 10.1074/jbc.M113.489336

    Figure Lengend Snippet: Mechanisms of Dsg2- and Dsg3-mediated cell-cell adhesion in keratinocytes. Targeting of Dsg3 binding either by pemphigus antibodies or siRNA leads to p38 MAPK activation and following retraction of the keratin filament network, which both negatively affect

    Article Snippet: Strikingly, 1 h of preincubation with the p38 MAPK inhibitor SB202190 (3.4 ± 1.2) or 24-h incubation of Dsg3-depleted cells with SB202190 significantly reduced the number of fragments (5.6 ± 1.5).

    Techniques: Binding Assay, Activation Assay

    Dsg3 depletion recruits Dsg2 and p-p38 MAPK to the plasma membrane, accompanied by alterations in PG protein and Dsg2 mRNA expression. A , representative Western blot analysis with detection of Dsg2 and PG in total cell lysates of MEK cells 24 h after

    Journal: The Journal of Biological Chemistry

    Article Title: Desmoglein 2 Compensates for Desmoglein 3 but Does Not Control Cell Adhesion via Regulation of p38 Mitogen-activated Protein Kinase in Keratinocytes

    doi: 10.1074/jbc.M113.489336

    Figure Lengend Snippet: Dsg3 depletion recruits Dsg2 and p-p38 MAPK to the plasma membrane, accompanied by alterations in PG protein and Dsg2 mRNA expression. A , representative Western blot analysis with detection of Dsg2 and PG in total cell lysates of MEK cells 24 h after

    Article Snippet: Strikingly, 1 h of preincubation with the p38 MAPK inhibitor SB202190 (3.4 ± 1.2) or 24-h incubation of Dsg3-depleted cells with SB202190 significantly reduced the number of fragments (5.6 ± 1.5).

    Techniques: Expressing, Western Blot