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    p38 MAP kinase inhibitor
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    99
    Millipore 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
    Sb202190, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2661 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 2661 article reviews
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    sb202190 - by Bioz Stars, 2021-01
    99/100 stars
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    95
    Tocris sb202190
    Inhibition of GSK3 promotes activation of the MKK3/6-p38 MAPK-ATF2 signaling module. ( a ) Immunoblot analysis for activating phosphorylations of proteins of the MKK3/6-p38 MAPK-ATF2 signaling module in primary brown adipocytes pre-treated with 10 μM SB216763 for 1 h before stimulation with 0.1 μM ISO for 1 h. ( b ) Immunoblot analysis of total and phosphorylated p38 MAPK in immortalized brown adipocytes overexpressing GSK3β mutants or empty vector. Cells were treated with 0.1 μM ISO for 1 h. ( c ) Fgf21 expression in primary brown adipocytes pre-treated with 10 μM <t>SB202190</t> (p38 MAPK inhibitor) for 1 h before treatment with 10 μM SB216763 (GSK3 inhibitor) for 1 h, followed by stimulation with 0.1 μM ISO for additional 6 h. ( d ) Schematic presentation of the proposed mechanism through which GSK3 regulates the thermogenic gene program in brown adipocytes. Data presented as mean of means +SEM (n = 3). Statistical significance was determined by two-way ANOVA with repeated measures and Tukey’s post hoc test for multiple comparisons. *p
    Sb202190, supplied by Tocris, used in various techniques. Bioz Stars score: 95/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 225 article reviews
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    95/100 stars
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    99
    Selleck Chemicals sb202190
    Differential expression of MAPK signaling-related genes in BHK-VECs. ( a ) qRT-PCR validation of DEGs related to MAPK pathways in BHK-VECs. GAPDH served as the internal reference gene to normalize data, and three biologically independent replicates were performed. ( b , c ) Effect of MAPK/ERK or <t>p38/MAPK</t> inhibition on replication of FMDV in BHK-21 cells. BHK-21 cells were pre-incubated (1 h) with DMSO, ( b ) 20 or 50 mM U0126, or ( c ) 20 or 50 mM <t>SB202190</t> and then infected with FMDV at 2.5 × 10 −4 PFU/cell for 24 h in the presence of DMSO, U0126, or SB202190. Protein extracts were examined using western blotting with FMDV 3D-specific or the indicated antibodies; the effectiveness of inhibition was monitored by detecting the phosphorylation of inhibitor-specific target protein (P-MAPK/ERK and pS15-Hsp27). ( d ) (i) p38δ (MAPK13) was down-regulated in BHK-VECs compared with BHK-21 cells, and (ii) overexpression of MAPK13 genes in BHK-21 cells promoted the replication of FMDV.
    Sb202190, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 99/100, based on 107 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 107 article reviews
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    sb202190 - by Bioz Stars, 2021-01
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    96
    Cell Signaling Technology Inc sb202190
    FSTL1 modulates myofibroblast differentiation by facilitating p38/JNK signaling. (a and b) Primary lung fibroblasts from Fstl1+/− and their WT littermates were treated with 5 ng/ml TGF-β1. (a) Immunofluorescence staining of α-SMA in lung fibroblasts. (b) Protein expression levels of α-SMA and type I collagen. (c-e) MLgs were pretreated with U0126, <t>SB202190,</t> and SP600125. Protein expression levels of α-SMA and type I collagen were detected by Western blotting. n = 4; Bars = 100 μm. * P
    Sb202190, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 96 stars, based on 122 article reviews
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    sb202190 - by Bioz Stars, 2021-01
    96/100 stars
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    95
    Selleck Chemicals sb202190 fhpi
    FSTL1 modulates myofibroblast differentiation by facilitating p38/JNK signaling. (a and b) Primary lung fibroblasts from Fstl1+/− and their WT littermates were treated with 5 ng/ml TGF-β1. (a) Immunofluorescence staining of α-SMA in lung fibroblasts. (b) Protein expression levels of α-SMA and type I collagen. (c-e) MLgs were pretreated with U0126, <t>SB202190,</t> and SP600125. Protein expression levels of α-SMA and type I collagen were detected by Western blotting. n = 4; Bars = 100 μm. * P
    Sb202190 Fhpi, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 95/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 8 article reviews
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    sb202190 fhpi - by Bioz Stars, 2021-01
    95/100 stars
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    94
    InvivoGen sb202190
    FSTL1 modulates myofibroblast differentiation by facilitating p38/JNK signaling. (a and b) Primary lung fibroblasts from Fstl1+/− and their WT littermates were treated with 5 ng/ml TGF-β1. (a) Immunofluorescence staining of α-SMA in lung fibroblasts. (b) Protein expression levels of α-SMA and type I collagen. (c-e) MLgs were pretreated with U0126, <t>SB202190,</t> and SP600125. Protein expression levels of α-SMA and type I collagen were detected by Western blotting. n = 4; Bars = 100 μm. * P
    Sb202190, supplied by InvivoGen, used in various techniques. Bioz Stars score: 94/100, based on 169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 169 article reviews
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    sb202190 - by Bioz Stars, 2021-01
    94/100 stars
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    92
    Merck KGaA sb202190
    NF-κB and C/EBPβ are required for induction of Tnfaip3 in LPS-activated macrophages. (A) Schematic map of predicted NF-κB p65 and C/EBPβ DNA binding sites in the promoter of Tnfaip3 . oPOSSUM ( http://www.cisreg.ca/oPOSSUM/ ) was used as the prediction tool. The JASPAR CORE vertebrate database was selected for transcription factor binding site matrices. Two arrows depict the PCR primers. (B) p65 and C/EBPβ bind to the promoter of Tnfaip3 after LPS stimulation. RAW264.7 cells were treated with LPS (100 ng/ml) for the indicated times. Chromatin was immunoprecipitated with anti-p65 and anti-C/EBPβ antibodies. Rabbit IgG was a negative control. Precipitated DNA or 1% of the chromatin input was amplified with primers for the Tnfaip3 promoter (−89 ∼ −410). The PCR products were loaded and separated on a 2% agarose gel. One of two independent experiments is shown. (C) LPS-induced association of p65 and C/EBPβ with Tnfaip3 was reduced in the presence of p38 inhibition. Chromatin isolated from RAW264.7 cells treated with LPS (100 ng/ml) for 4 h in the absence or presence of <t>SB202190</t> (10 µM) were subjected to ChIP assay as described above. The relative quantity of promoter enriched by ChIP was quantified by real-time PCR and expressed as the fold enrichment of untreated control samples after normalization to rabbit IgG. Data represent the means of two independent experiments. (D) LPS-induced expression of A20 (TNFAIP3) was decreased in C/EBPβ-depleted RAW264.7 cells. Cells were infected with lentiviruses encoding shRNA against luciferase ( shLuc ) or C/EBPβ ( shCebpb ), and treated with LPS (100 ng/ml) for the indicated times. Cell lysates were collected and analyzed by immunoblotting using the indicated antibodies.
    Sb202190, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sb202190/product/Merck KGaA
    Average 92 stars, based on 33 article reviews
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    sb202190 - by Bioz Stars, 2021-01
    92/100 stars
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    92
    ApexBio sb202190
    Effect of PD98059 (PD), a p44/42 inhibitor, <t>SB202190</t> (SB), a p38 inhibitor, and SP600125 (SP), a JNK inhibitor, on PAK4 phosphorylation ( A ) and on CREB phosphorylation ( B ) by secretin and VIP in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of PD98059 (10 µM), SB202190 (10 µM) or SP600125 (20 µM) for 1 h and then incubated with no addition (control), secretin (10 nM) of VIP (10 nM) for 15 min and then lysed. Western blots were analyzed using anti-pS474 PAK4 or pS133 CREB and, as loading control, antitotal PAK4 or antitotal CREB. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of stimulation over the control group. * P
    Sb202190, supplied by ApexBio, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sb202190/product/ApexBio
    Average 92 stars, based on 23 article reviews
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    sb202190 - by Bioz Stars, 2021-01
    92/100 stars
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    92
    Biomol GmbH sb202190
    Effects of garcinol and p300 siRNA on leptin-stimulated A549 cells. Serum-starved A549 cells were pretreated with the OB-R antibody (2 µg/mL), U0126 (10 µM), <t>SB202190</t> (10 µM), SP600125 (10 µM), Bay11-7082 (10 µM), or garcinol (1 µM) for 1 h. Other cells were transfected with 100 nM p300 siRNA or scrambled siRNA as described in the Experimental section. The cells were then incubated with 1 µg/mL of leptin for the indicated time intervals. At the end of the incubation, the cells were harvested and cell lysates were extracted. ( A , B ) Expression of cPLA 2 -α and p300; ( C ) phosphorylation of p65; and ( D ) acetylation of histone H4 were detected using Western blot with specific antibodies. Cell membranes were stripped and reprobed with the anti-GAPDH antibody as internal controls. The data are expressed as mean ± SEM of 5 independent experiments ( n = 5). p
    Sb202190, supplied by Biomol GmbH, used in various techniques. Bioz Stars score: 92/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 96 article reviews
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    sb202190 - by Bioz Stars, 2021-01
    92/100 stars
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    93
    Cayman Chemical sb202190
    TLR2-MAPK-ERK1/2 signaling cascade is required for the induction of VEGFA and Snail-1 by PCN033 ( A ) Phosphorylation of ERK1/2 along with PCN033 infection. The β-actin was detected as the loading control. ( B ) Densitometrical analysis of the ERK1/2 activation in hBMEC 2 h post-infection, compared with that in uninfected cells. Data are calculated as the ratio of phospho-ERK1/2 to total ERK1/2. ( C ) Effects of the MAPK signaling inhibitors on the PCN033-induced upregulation of VEGFA. U0126 (selective inhibitor of ERK1/2) and <t>SB202190</t> (selective inhibitor of p38) could significantly decrease the PCN033-induced upregulation of VEGFA, while SP600125 (specific inhibitor of JNK) could not. ( D ) Effects of the MAPK signaling inhibitors on the PCN033-induced upregulation of Snail-1. Selective ERK1/2 inhibitor U0126 could completely block the PCN033-induced upregulation of Snail-1. ( E – F ) Snail-1 knocking-down via shRNA in hBMEC did not affect the induction of VEGFA by PCN033, while blocking VEGFA pathway significantly decreased the upregulation of Snail-1. ( G – H ) Effects of the VEGFR inhibitors on PCN033-induced activation of ERK1/2 and the densitometric analysis. ( I – J ) TLR2 agonist Pam3CSK4 induced the activation of ERK1/2 in a dose-dependent manner. ( K – L ) Pam3CSK4 dose-dependently induced the upregulation of Snail-1 and VEGFA in hBMEC. Results are expressed as mean ± SD from three independent assays.
    Sb202190, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 29 article reviews
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    sb202190 - by Bioz Stars, 2021-01
    93/100 stars
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    92
    LC Laboratories sb202190
    Suppression ­­­of p38 signaling restores sensitivity to PIM inhibition in primary AML cells and mouse xenografts A.-B. Inhibition of p38 enhances AZD1208 response in primary AML cells. Patient cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.313 μM, 1.25 μM, or 5 μM SCIO-469. Growth was measured after 4 days using MTT. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). C. Combined p38/PIM inhibition suppresses mTOR signaling. Primary AML cells were treated for 24 hours with 1 μM AZD1208, 10 μM SCIO-469, or the combination. Cell lysates were harvested and subjected to western blot analysis. D.-G. Inhibition of p38 enhances AZD1208 response in a validation set of primary AML cells. Cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.3 μM, 1 μM, or 3 μM SCIO-469. Viability was measured after 72 hours using CellTiter-Glo. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). H. SCIO-469 is synergistic with AZD1208. Primary AML cells were treated with 2-fold dilutions of AZD1208, SCIO-469, or the combination. Viability was assessed by CellTiter-Blue and used to calculate synergy scores. I. Dual PIM/p38 inhibition suppresses tumor growth in a xenograft model. K562 cells (5.10 6 ) were subcutaneously implanted in Rag2 −/− IL2γc −/− mice. Once tumors were established, animals were treated with vehicle, AZD1208 (30 mg/kg), <t>SB202190</t> (5 mg/kg), or both drugs in combination. J. Mean tumor volume after 18 days treatment. Vehicle ( n = 11), AZD1208 ( n = 12), SB202190 ( n = 18), and combination ( n = 14). P -values were calculated via one-way ANOVA and Dunnett's test.
    Sb202190, supplied by LC Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Enzo Biochem sb202190
    Suppression ­­­of p38 signaling restores sensitivity to PIM inhibition in primary AML cells and mouse xenografts A.-B. Inhibition of p38 enhances AZD1208 response in primary AML cells. Patient cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.313 μM, 1.25 μM, or 5 μM SCIO-469. Growth was measured after 4 days using MTT. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). C. Combined p38/PIM inhibition suppresses mTOR signaling. Primary AML cells were treated for 24 hours with 1 μM AZD1208, 10 μM SCIO-469, or the combination. Cell lysates were harvested and subjected to western blot analysis. D.-G. Inhibition of p38 enhances AZD1208 response in a validation set of primary AML cells. Cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.3 μM, 1 μM, or 3 μM SCIO-469. Viability was measured after 72 hours using CellTiter-Glo. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). H. SCIO-469 is synergistic with AZD1208. Primary AML cells were treated with 2-fold dilutions of AZD1208, SCIO-469, or the combination. Viability was assessed by CellTiter-Blue and used to calculate synergy scores. I. Dual PIM/p38 inhibition suppresses tumor growth in a xenograft model. K562 cells (5.10 6 ) were subcutaneously implanted in Rag2 −/− IL2γc −/− mice. Once tumors were established, animals were treated with vehicle, AZD1208 (30 mg/kg), <t>SB202190</t> (5 mg/kg), or both drugs in combination. J. Mean tumor volume after 18 days treatment. Vehicle ( n = 11), AZD1208 ( n = 12), SB202190 ( n = 18), and combination ( n = 14). P -values were calculated via one-way ANOVA and Dunnett's test.
    Sb202190, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 93/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 25 article reviews
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    sb202190 - by Bioz Stars, 2021-01
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    93
    Abcam sb202190
    Phospho-p38 MAPK inhibitors prevent and reverse EndoMT-associated gene expression in vitro. ( a ) To study possible prevention of EndoMT, hREC were treated with the p38 MAPK inhibitor SB203580 or <t>SB202190</t> (10 μM) during EndoMT induction. Expression levels for the genes encoding the EndoMT-associated proteins SNAI1, SNAI2, α-SMA, FSP-1, vimentin, fibronectin, COL1A2 and COL3A1; and the endothelial differentiation markers VE-cadherin, CD31 and VEGFR2 were assessed by qPCR on day 6 of EndoMT induction. Data = mean ± SEM, * p
    Sb202190, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 12 article reviews
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    sb202190 - by Bioz Stars, 2021-01
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    N/A
    SB202190 hydrochloride is a water soluble form of the potent p38 SAPK2a inhibitor SB202190 sc 202334
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    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 chromatin immunoprecipitation (ChIP) assays, HUVEC were stimulated with 10 ng/ml TNF-α (R & D Systems, Inc.) for 10 min. For p38 inhibitor ChIP assays, HUVEC were treated for 2 h with 20 μM p38 inhibitor SB202190 (Sigma Aldrich) or DMSO (Carl Roth GmbH & Co., KG) as solvent control prior to 10 ng/ml TNF-α treatment (R & D Systems, Inc.) for 10 min.

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

    Inhibition of phosphorylation of MAPK blocks CAPE-enhanced expression of NDRG1 in TW01 cells. ( A )The expression levels of ERK, p-ERK, NDRG1, or β-actin after pretreatments with (+) or without (−) PD0325901 for 1 h before 30 μM CAPE treatment in TW01 cells were determined by immunoblotting and RT-qPCR ( B ) assays. TW01 cells were pretreated with (+) or without (−) SP600125 ( C ) or SB202190 ( D ). The protein levels of JNK, p-JNK, p38, p-p38, NDRG1, or β-actin in CAPE-treated TW01 cells were determined by immunoblotting assay. (** p

    Journal: International Journal of Molecular Sciences

    Article Title: Caffeic Acid Phenethyl Ester Induces N-myc Downstream Regulated Gene 1 to Inhibit Cell Proliferation and Invasion of Human Nasopharyngeal Cancer Cells

    doi: 10.3390/ijms19051397

    Figure Lengend Snippet: Inhibition of phosphorylation of MAPK blocks CAPE-enhanced expression of NDRG1 in TW01 cells. ( A )The expression levels of ERK, p-ERK, NDRG1, or β-actin after pretreatments with (+) or without (−) PD0325901 for 1 h before 30 μM CAPE treatment in TW01 cells were determined by immunoblotting and RT-qPCR ( B ) assays. TW01 cells were pretreated with (+) or without (−) SP600125 ( C ) or SB202190 ( D ). The protein levels of JNK, p-JNK, p38, p-p38, NDRG1, or β-actin in CAPE-treated TW01 cells were determined by immunoblotting assay. (** p

    Article Snippet: CAPE, p38 inhibitor (SB202190), and ERK inhibitor (PD0325901) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

    Techniques: Inhibition, Expressing, Quantitative RT-PCR

    CAPE modulates phosphorylation of MAPK and STAT3 in NPC cells. ( A ) Time course of activities of components of MAPK signal pathway and STAT3 in TW04 cells treated with 30 μM CAPE. Expressions of ERK, p-ERK, p38, p-p38, JNK, p-JNK, STAT3, p-STAT3, and β-actin were determined by the immunoblotting assay. ( B ) Dose responses of ERK, p-ERK, p38, p-p38, JNK, p-JNK, STAT3, p-STAT3, and β-actin were determined by the immunoblotting assay after 15 min of CAPE treatments in TW04 cells. ( C ) Time course of activities of components of MAPK signal pathway and STAT3 in TW01 cells treated with 30 μM CAPE were evaluated by immunoblotting assay. ( D ) The pSTAT3-TA-Luc reporter vectors were transfected into TW04 cells for 24 h, and cells were then treated by indicated concentrations of CAPE for 24 h. Data were presented as the mean percentage of luciferase activity induced by the CAPE treatment relative to the control solvent-treated group (±SE, n = 6). (* p

    Journal: International Journal of Molecular Sciences

    Article Title: Caffeic Acid Phenethyl Ester Induces N-myc Downstream Regulated Gene 1 to Inhibit Cell Proliferation and Invasion of Human Nasopharyngeal Cancer Cells

    doi: 10.3390/ijms19051397

    Figure Lengend Snippet: CAPE modulates phosphorylation of MAPK and STAT3 in NPC cells. ( A ) Time course of activities of components of MAPK signal pathway and STAT3 in TW04 cells treated with 30 μM CAPE. Expressions of ERK, p-ERK, p38, p-p38, JNK, p-JNK, STAT3, p-STAT3, and β-actin were determined by the immunoblotting assay. ( B ) Dose responses of ERK, p-ERK, p38, p-p38, JNK, p-JNK, STAT3, p-STAT3, and β-actin were determined by the immunoblotting assay after 15 min of CAPE treatments in TW04 cells. ( C ) Time course of activities of components of MAPK signal pathway and STAT3 in TW01 cells treated with 30 μM CAPE were evaluated by immunoblotting assay. ( D ) The pSTAT3-TA-Luc reporter vectors were transfected into TW04 cells for 24 h, and cells were then treated by indicated concentrations of CAPE for 24 h. Data were presented as the mean percentage of luciferase activity induced by the CAPE treatment relative to the control solvent-treated group (±SE, n = 6). (* p

    Article Snippet: CAPE, p38 inhibitor (SB202190), and ERK inhibitor (PD0325901) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

    Techniques: Transfection, Luciferase, Activity Assay

    Regulation of IGFBP-4 expression in M21 cells involves altered p38 MAPK signaling. M21 melanoma cells expressing (M21) or lacking (M21L) αvβ3 were examined for the expression of total and activated p38 MAPK and Erk. a Western blots of

    Journal: Angiogenesis

    Article Title: Inhibition of tumor-associated αvβ3 integrin regulates the angiogenic switch by enhancing expression of IGFBP-4 leading to reduced melanoma growth and angiogenesis in vivo

    doi: 10.1007/s10456-014-9445-2

    Figure Lengend Snippet: Regulation of IGFBP-4 expression in M21 cells involves altered p38 MAPK signaling. M21 melanoma cells expressing (M21) or lacking (M21L) αvβ3 were examined for the expression of total and activated p38 MAPK and Erk. a Western blots of

    Article Snippet: P38 MAPK inhibitor (SB202190) was obtained from EMD chemicals (Gibbston, NJ).

    Techniques: Expressing, Western Blot

    Inhibition of GSK3 promotes activation of the MKK3/6-p38 MAPK-ATF2 signaling module. ( a ) Immunoblot analysis for activating phosphorylations of proteins of the MKK3/6-p38 MAPK-ATF2 signaling module in primary brown adipocytes pre-treated with 10 μM SB216763 for 1 h before stimulation with 0.1 μM ISO for 1 h. ( b ) Immunoblot analysis of total and phosphorylated p38 MAPK in immortalized brown adipocytes overexpressing GSK3β mutants or empty vector. Cells were treated with 0.1 μM ISO for 1 h. ( c ) Fgf21 expression in primary brown adipocytes pre-treated with 10 μM SB202190 (p38 MAPK inhibitor) for 1 h before treatment with 10 μM SB216763 (GSK3 inhibitor) for 1 h, followed by stimulation with 0.1 μM ISO for additional 6 h. ( d ) Schematic presentation of the proposed mechanism through which GSK3 regulates the thermogenic gene program in brown adipocytes. Data presented as mean of means +SEM (n = 3). Statistical significance was determined by two-way ANOVA with repeated measures and Tukey’s post hoc test for multiple comparisons. *p

    Journal: Scientific Reports

    Article Title: GSK3 is a negative regulator of the thermogenic program in brown adipocytes

    doi: 10.1038/s41598-018-21795-y

    Figure Lengend Snippet: Inhibition of GSK3 promotes activation of the MKK3/6-p38 MAPK-ATF2 signaling module. ( a ) Immunoblot analysis for activating phosphorylations of proteins of the MKK3/6-p38 MAPK-ATF2 signaling module in primary brown adipocytes pre-treated with 10 μM SB216763 for 1 h before stimulation with 0.1 μM ISO for 1 h. ( b ) Immunoblot analysis of total and phosphorylated p38 MAPK in immortalized brown adipocytes overexpressing GSK3β mutants or empty vector. Cells were treated with 0.1 μM ISO for 1 h. ( c ) Fgf21 expression in primary brown adipocytes pre-treated with 10 μM SB202190 (p38 MAPK inhibitor) for 1 h before treatment with 10 μM SB216763 (GSK3 inhibitor) for 1 h, followed by stimulation with 0.1 μM ISO for additional 6 h. ( d ) Schematic presentation of the proposed mechanism through which GSK3 regulates the thermogenic gene program in brown adipocytes. Data presented as mean of means +SEM (n = 3). Statistical significance was determined by two-way ANOVA with repeated measures and Tukey’s post hoc test for multiple comparisons. *p

    Article Snippet: SB415286, SB216763, BIO, H89 and SB202190 were obtained from Tocris Bioscience.

    Techniques: Inhibition, Activation Assay, Plasmid Preparation, Expressing

    Differential expression of MAPK signaling-related genes in BHK-VECs. ( a ) qRT-PCR validation of DEGs related to MAPK pathways in BHK-VECs. GAPDH served as the internal reference gene to normalize data, and three biologically independent replicates were performed. ( b , c ) Effect of MAPK/ERK or p38/MAPK inhibition on replication of FMDV in BHK-21 cells. BHK-21 cells were pre-incubated (1 h) with DMSO, ( b ) 20 or 50 mM U0126, or ( c ) 20 or 50 mM SB202190 and then infected with FMDV at 2.5 × 10 −4 PFU/cell for 24 h in the presence of DMSO, U0126, or SB202190. Protein extracts were examined using western blotting with FMDV 3D-specific or the indicated antibodies; the effectiveness of inhibition was monitored by detecting the phosphorylation of inhibitor-specific target protein (P-MAPK/ERK and pS15-Hsp27). ( d ) (i) p38δ (MAPK13) was down-regulated in BHK-VECs compared with BHK-21 cells, and (ii) overexpression of MAPK13 genes in BHK-21 cells promoted the replication of FMDV.

    Journal: Scientific Reports

    Article Title: Cellular response to persistent foot-and-mouth disease virus infection is linked to specific types of alterations in the host cell transcriptome

    doi: 10.1038/s41598-018-23478-0

    Figure Lengend Snippet: Differential expression of MAPK signaling-related genes in BHK-VECs. ( a ) qRT-PCR validation of DEGs related to MAPK pathways in BHK-VECs. GAPDH served as the internal reference gene to normalize data, and three biologically independent replicates were performed. ( b , c ) Effect of MAPK/ERK or p38/MAPK inhibition on replication of FMDV in BHK-21 cells. BHK-21 cells were pre-incubated (1 h) with DMSO, ( b ) 20 or 50 mM U0126, or ( c ) 20 or 50 mM SB202190 and then infected with FMDV at 2.5 × 10 −4 PFU/cell for 24 h in the presence of DMSO, U0126, or SB202190. Protein extracts were examined using western blotting with FMDV 3D-specific or the indicated antibodies; the effectiveness of inhibition was monitored by detecting the phosphorylation of inhibitor-specific target protein (P-MAPK/ERK and pS15-Hsp27). ( d ) (i) p38δ (MAPK13) was down-regulated in BHK-VECs compared with BHK-21 cells, and (ii) overexpression of MAPK13 genes in BHK-21 cells promoted the replication of FMDV.

    Article Snippet: MEK1/2 inhibitor U0126 and P38 inhibitor SB202190 were purchased from Selleck.

    Techniques: Expressing, Quantitative RT-PCR, Inhibition, Incubation, Infection, Western Blot, Over Expression

    FSTL1 modulates myofibroblast differentiation by facilitating p38/JNK signaling. (a and b) Primary lung fibroblasts from Fstl1+/− and their WT littermates were treated with 5 ng/ml TGF-β1. (a) Immunofluorescence staining of α-SMA in lung fibroblasts. (b) Protein expression levels of α-SMA and type I collagen. (c-e) MLgs were pretreated with U0126, SB202190, and SP600125. Protein expression levels of α-SMA and type I collagen were detected by Western blotting. n = 4; Bars = 100 μm. * P

    Journal: Chinese Medical Journal

    Article Title: Follistatin-Like 1 Promotes Bleomycin-Induced Pulmonary Fibrosis through the Transforming Growth Factor Beta 1/Mitogen-Activated Protein Kinase Signaling Pathway

    doi: 10.4103/0366-6999.238151

    Figure Lengend Snippet: FSTL1 modulates myofibroblast differentiation by facilitating p38/JNK signaling. (a and b) Primary lung fibroblasts from Fstl1+/− and their WT littermates were treated with 5 ng/ml TGF-β1. (a) Immunofluorescence staining of α-SMA in lung fibroblasts. (b) Protein expression levels of α-SMA and type I collagen. (c-e) MLgs were pretreated with U0126, SB202190, and SP600125. Protein expression levels of α-SMA and type I collagen were detected by Western blotting. n = 4; Bars = 100 μm. * P

    Article Snippet: MLgs were cultured in 96-well plates for 24 h, and serum starved for 24 h. The cells were preincubated for 1 h with U0126 (ERK inhibitor, Cell Signaling Technology, MA, USA), SB202190 (p38 inhibitor, Cell Signaling Technology, MA, USA), SP600125 (JNK inhibitor, Cell Signaling Technology, MA, USA), and SB525334 (Smad2/3 inhibitor, R and D Systems, MN, USA) before treatment with 5 ng/ml TGF-β1 (R and D Systems, MN, USA) in the presence or absence of 100 ng/ml recombinant human FSTL1 protein (R and D Systems, MN, USA) for 24 h and finally incubated with MTT (final concentration of 0.5 mg/ml) for 4 h. The supernatant was removed, and 150 ml/well of dimethylsulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) was added to dissolve the blue formazan crystals by shaking the plates for 15 min on an orbital shaker at 25°C.

    Techniques: Immunofluorescence, Staining, Expressing, Western Blot

    Effect of inhibition of the p38 signaling pathway on cell migration and the protein expression of MMPs in astrocytes of the untreated, SB202190 and (DES) + SB202190 groups. (A) Cell migration was determined by a Transwell migration assay. (B) Representative western blot bands for the protein expression of MMP-2 and MMP-9 in the untreated, SB202190 and DES + SB202190 groups. (C) Densitometric analysis was performed to obtain the relative expression level of target proteins with β-actin as the internal reference. SB202190 was employed as an inhibitor of the p38 signaling pathway, while DES was employed as an activator of protein phosphatase 2A. **P

    Journal: Molecular Medicine Reports

    Article Title: Protein phosphatase 2A regulates the p38 signaling pathway to affect the migration of astrocytes

    doi: 10.3892/mmr.2018.9425

    Figure Lengend Snippet: Effect of inhibition of the p38 signaling pathway on cell migration and the protein expression of MMPs in astrocytes of the untreated, SB202190 and (DES) + SB202190 groups. (A) Cell migration was determined by a Transwell migration assay. (B) Representative western blot bands for the protein expression of MMP-2 and MMP-9 in the untreated, SB202190 and DES + SB202190 groups. (C) Densitometric analysis was performed to obtain the relative expression level of target proteins with β-actin as the internal reference. SB202190 was employed as an inhibitor of the p38 signaling pathway, while DES was employed as an activator of protein phosphatase 2A. **P

    Article Snippet: Cells in the SB202190 group were incubated at 37°C for 48 h with cell culture medium containing the p38 signal pathway inhibitor SB202190 (Cell Signaling Technology, Inc.) at a final concentration of 30 µmol/l.

    Techniques: Inhibition, Migration, Expressing, Transwell Migration Assay, Western Blot

    Effect of a p38 signaling pathway inhibitor on p-p38 protein expression in astrocytes. (A) Representative western blot bands for the protein expression of p-p38 and p38 in untreated astrocytes and astrocytes treated with the p38 signaling pathway inhibitor, SB202190. (B) Densitometric analysis was performed to obtain the relative expression level of p-p38 protein with p38 as the internal reference. **P

    Journal: Molecular Medicine Reports

    Article Title: Protein phosphatase 2A regulates the p38 signaling pathway to affect the migration of astrocytes

    doi: 10.3892/mmr.2018.9425

    Figure Lengend Snippet: Effect of a p38 signaling pathway inhibitor on p-p38 protein expression in astrocytes. (A) Representative western blot bands for the protein expression of p-p38 and p38 in untreated astrocytes and astrocytes treated with the p38 signaling pathway inhibitor, SB202190. (B) Densitometric analysis was performed to obtain the relative expression level of p-p38 protein with p38 as the internal reference. **P

    Article Snippet: Cells in the SB202190 group were incubated at 37°C for 48 h with cell culture medium containing the p38 signal pathway inhibitor SB202190 (Cell Signaling Technology, Inc.) at a final concentration of 30 µmol/l.

    Techniques: Expressing, Western Blot

    NF-κB and C/EBPβ are required for induction of Tnfaip3 in LPS-activated macrophages. (A) Schematic map of predicted NF-κB p65 and C/EBPβ DNA binding sites in the promoter of Tnfaip3 . oPOSSUM ( http://www.cisreg.ca/oPOSSUM/ ) was used as the prediction tool. The JASPAR CORE vertebrate database was selected for transcription factor binding site matrices. Two arrows depict the PCR primers. (B) p65 and C/EBPβ bind to the promoter of Tnfaip3 after LPS stimulation. RAW264.7 cells were treated with LPS (100 ng/ml) for the indicated times. Chromatin was immunoprecipitated with anti-p65 and anti-C/EBPβ antibodies. Rabbit IgG was a negative control. Precipitated DNA or 1% of the chromatin input was amplified with primers for the Tnfaip3 promoter (−89 ∼ −410). The PCR products were loaded and separated on a 2% agarose gel. One of two independent experiments is shown. (C) LPS-induced association of p65 and C/EBPβ with Tnfaip3 was reduced in the presence of p38 inhibition. Chromatin isolated from RAW264.7 cells treated with LPS (100 ng/ml) for 4 h in the absence or presence of SB202190 (10 µM) were subjected to ChIP assay as described above. The relative quantity of promoter enriched by ChIP was quantified by real-time PCR and expressed as the fold enrichment of untreated control samples after normalization to rabbit IgG. Data represent the means of two independent experiments. (D) LPS-induced expression of A20 (TNFAIP3) was decreased in C/EBPβ-depleted RAW264.7 cells. Cells were infected with lentiviruses encoding shRNA against luciferase ( shLuc ) or C/EBPβ ( shCebpb ), and treated with LPS (100 ng/ml) for the indicated times. Cell lysates were collected and analyzed by immunoblotting using the indicated antibodies.

    Journal: PLoS ONE

    Article Title: Transcription of Tnfaip3 Is Regulated by NF-?B and p38 via C/EBP? in Activated Macrophages

    doi: 10.1371/journal.pone.0073153

    Figure Lengend Snippet: NF-κB and C/EBPβ are required for induction of Tnfaip3 in LPS-activated macrophages. (A) Schematic map of predicted NF-κB p65 and C/EBPβ DNA binding sites in the promoter of Tnfaip3 . oPOSSUM ( http://www.cisreg.ca/oPOSSUM/ ) was used as the prediction tool. The JASPAR CORE vertebrate database was selected for transcription factor binding site matrices. Two arrows depict the PCR primers. (B) p65 and C/EBPβ bind to the promoter of Tnfaip3 after LPS stimulation. RAW264.7 cells were treated with LPS (100 ng/ml) for the indicated times. Chromatin was immunoprecipitated with anti-p65 and anti-C/EBPβ antibodies. Rabbit IgG was a negative control. Precipitated DNA or 1% of the chromatin input was amplified with primers for the Tnfaip3 promoter (−89 ∼ −410). The PCR products were loaded and separated on a 2% agarose gel. One of two independent experiments is shown. (C) LPS-induced association of p65 and C/EBPβ with Tnfaip3 was reduced in the presence of p38 inhibition. Chromatin isolated from RAW264.7 cells treated with LPS (100 ng/ml) for 4 h in the absence or presence of SB202190 (10 µM) were subjected to ChIP assay as described above. The relative quantity of promoter enriched by ChIP was quantified by real-time PCR and expressed as the fold enrichment of untreated control samples after normalization to rabbit IgG. Data represent the means of two independent experiments. (D) LPS-induced expression of A20 (TNFAIP3) was decreased in C/EBPβ-depleted RAW264.7 cells. Cells were infected with lentiviruses encoding shRNA against luciferase ( shLuc ) or C/EBPβ ( shCebpb ), and treated with LPS (100 ng/ml) for the indicated times. Cell lysates were collected and analyzed by immunoblotting using the indicated antibodies.

    Article Snippet: For inhibition of p38, BMDMs from C57BL/6 mice were treated with 10 µM SB202190 (Merck, Germany) for 2 h prior to use.

    Techniques: Binding Assay, Polymerase Chain Reaction, Immunoprecipitation, Negative Control, Amplification, Agarose Gel Electrophoresis, Inhibition, Isolation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Expressing, Infection, shRNA, Luciferase

    Depletion of IKKβ expression and inhibition of p38 signaling pathway in Ikkβ Δ and SB202190-treated bone marrow-derived macrophages (BMDMs). (A) Immunoblotting of IKKβ and p38 from BMDMs isolated from wild-type (wt; Ikkβ F/F ) and Ikkβ Δ mice. (B) Immunoblotting of p38 and phosphorylated p38 from wt BMDMs treated with LPS (100 ng/mL) in the absence or presence of SB202190 (10 µM) for 2 h. (C D) mRNA expression levels of IL-1β and IL-6 were inhibited in SB202190-treated BMDMs after LPS treatment. The expression levels of Il1b (C) and Il6 (D) were determined from wt BMDMs treated with LPS (100 ng/mL) for 4 h in the absence or presence of SB202190 (10 µM) for 2 h using real-time RT-PCR. Data represent the mean ± SEM for three independent experiments. ***, P

    Journal: PLoS ONE

    Article Title: Transcription of Tnfaip3 Is Regulated by NF-?B and p38 via C/EBP? in Activated Macrophages

    doi: 10.1371/journal.pone.0073153

    Figure Lengend Snippet: Depletion of IKKβ expression and inhibition of p38 signaling pathway in Ikkβ Δ and SB202190-treated bone marrow-derived macrophages (BMDMs). (A) Immunoblotting of IKKβ and p38 from BMDMs isolated from wild-type (wt; Ikkβ F/F ) and Ikkβ Δ mice. (B) Immunoblotting of p38 and phosphorylated p38 from wt BMDMs treated with LPS (100 ng/mL) in the absence or presence of SB202190 (10 µM) for 2 h. (C D) mRNA expression levels of IL-1β and IL-6 were inhibited in SB202190-treated BMDMs after LPS treatment. The expression levels of Il1b (C) and Il6 (D) were determined from wt BMDMs treated with LPS (100 ng/mL) for 4 h in the absence or presence of SB202190 (10 µM) for 2 h using real-time RT-PCR. Data represent the mean ± SEM for three independent experiments. ***, P

    Article Snippet: For inhibition of p38, BMDMs from C57BL/6 mice were treated with 10 µM SB202190 (Merck, Germany) for 2 h prior to use.

    Techniques: Expressing, Inhibition, Derivative Assay, Isolation, Mouse Assay, Quantitative RT-PCR

    Identification of LPS-induced genes that were regulated by both NF-κB and p38. (A) Venn diagram of NF-κB and p38-dependent genes. NF-κB-related genes were identified from genes that were down-regulated in Ikkβ Δ BMDMs as compared with wt BMDMs after LPS treatment, and p38-related genes were selected by comparing SB202190- (p38-inhibitor) with dimethyl sulfoxide (DMSO)-treated BMDMs after LPS treatment. Thirty-two LPS-induced genes were regulated by both NF-κB and p38-downstream transcription factors. (B) Hierarchical clustering of average fold change for the NF-κB and p38-dependent genes. Each column represents the average fold change 4 h after LPS treatment compared to 0 h. *: genes chosen for PCR validation. (C) Relative fold changes of Il1b, Serpinb2, Tnfaip3 , and Zc3h12a mRNA from BMDMs from wt and Ikkβ Δ cells stimulated with LPS (100 ng/mL) for 4 h in the presence or absence of SB202190 (10 µM) were measured by quantitative RT-PCR. The fold change of 4 h vs. 0 h was normalized to wt BMDMs. The internal control was Cyclophilin A mRNA ( Cypa ). Data represent the mean ± SD for at least two independent experiments. *, P

    Journal: PLoS ONE

    Article Title: Transcription of Tnfaip3 Is Regulated by NF-?B and p38 via C/EBP? in Activated Macrophages

    doi: 10.1371/journal.pone.0073153

    Figure Lengend Snippet: Identification of LPS-induced genes that were regulated by both NF-κB and p38. (A) Venn diagram of NF-κB and p38-dependent genes. NF-κB-related genes were identified from genes that were down-regulated in Ikkβ Δ BMDMs as compared with wt BMDMs after LPS treatment, and p38-related genes were selected by comparing SB202190- (p38-inhibitor) with dimethyl sulfoxide (DMSO)-treated BMDMs after LPS treatment. Thirty-two LPS-induced genes were regulated by both NF-κB and p38-downstream transcription factors. (B) Hierarchical clustering of average fold change for the NF-κB and p38-dependent genes. Each column represents the average fold change 4 h after LPS treatment compared to 0 h. *: genes chosen for PCR validation. (C) Relative fold changes of Il1b, Serpinb2, Tnfaip3 , and Zc3h12a mRNA from BMDMs from wt and Ikkβ Δ cells stimulated with LPS (100 ng/mL) for 4 h in the presence or absence of SB202190 (10 µM) were measured by quantitative RT-PCR. The fold change of 4 h vs. 0 h was normalized to wt BMDMs. The internal control was Cyclophilin A mRNA ( Cypa ). Data represent the mean ± SD for at least two independent experiments. *, P

    Article Snippet: For inhibition of p38, BMDMs from C57BL/6 mice were treated with 10 µM SB202190 (Merck, Germany) for 2 h prior to use.

    Techniques: Polymerase Chain Reaction, Quantitative RT-PCR

    C/EBPβ and A20 (TNFAIP3) were suppressed in Ikkβ Δ and p38-inhibited macrophages. (A–C) Expression levels of Tnfaip3 and Cebpb mRNA were decreased in Ikkβ Δ and p38-inhibited BMDMs in response to LPS. BMDMs from wt and Ikkβ Δ cells treated with or without SB202190 (10 µM) were stimulated with LPS (100 ng/mL) for 4 h. Total RNAs were isolated and analyzed by semi-quantitative RT-PCR (A) or quantitative real-time RT-PCR for expression of Cebpb (B) and Tnfaip3 (C) mRNAs. Results were normalized to Cyclophilin A ( Cypa ) and are presented relative to expression in wt BMDMs. * P

    Journal: PLoS ONE

    Article Title: Transcription of Tnfaip3 Is Regulated by NF-?B and p38 via C/EBP? in Activated Macrophages

    doi: 10.1371/journal.pone.0073153

    Figure Lengend Snippet: C/EBPβ and A20 (TNFAIP3) were suppressed in Ikkβ Δ and p38-inhibited macrophages. (A–C) Expression levels of Tnfaip3 and Cebpb mRNA were decreased in Ikkβ Δ and p38-inhibited BMDMs in response to LPS. BMDMs from wt and Ikkβ Δ cells treated with or without SB202190 (10 µM) were stimulated with LPS (100 ng/mL) for 4 h. Total RNAs were isolated and analyzed by semi-quantitative RT-PCR (A) or quantitative real-time RT-PCR for expression of Cebpb (B) and Tnfaip3 (C) mRNAs. Results were normalized to Cyclophilin A ( Cypa ) and are presented relative to expression in wt BMDMs. * P

    Article Snippet: For inhibition of p38, BMDMs from C57BL/6 mice were treated with 10 µM SB202190 (Merck, Germany) for 2 h prior to use.

    Techniques: Expressing, Isolation, Quantitative RT-PCR

    Effect of PD98059 (PD), a p44/42 inhibitor, SB202190 (SB), a p38 inhibitor, and SP600125 (SP), a JNK inhibitor, on PAK4 phosphorylation ( A ) and on CREB phosphorylation ( B ) by secretin and VIP in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of PD98059 (10 µM), SB202190 (10 µM) or SP600125 (20 µM) for 1 h and then incubated with no addition (control), secretin (10 nM) of VIP (10 nM) for 15 min and then lysed. Western blots were analyzed using anti-pS474 PAK4 or pS133 CREB and, as loading control, antitotal PAK4 or antitotal CREB. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of stimulation over the control group. * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Hormones, Neurotransmitters, Growth Factors, Receptors, and Signaling: Cyclic AMP-dependent protein kinase A and EPAC mediate VIP and secretin stimulation of PAK4 and activation of Na+,K+-ATPase in pancreatic acinar cells

    doi: 10.1152/ajpgi.00275.2018

    Figure Lengend Snippet: Effect of PD98059 (PD), a p44/42 inhibitor, SB202190 (SB), a p38 inhibitor, and SP600125 (SP), a JNK inhibitor, on PAK4 phosphorylation ( A ) and on CREB phosphorylation ( B ) by secretin and VIP in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of PD98059 (10 µM), SB202190 (10 µM) or SP600125 (20 µM) for 1 h and then incubated with no addition (control), secretin (10 nM) of VIP (10 nM) for 15 min and then lysed. Western blots were analyzed using anti-pS474 PAK4 or pS133 CREB and, as loading control, antitotal PAK4 or antitotal CREB. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of stimulation over the control group. * P

    Article Snippet: PD98059 and SB202190 were from APExBIO (Houston, TX).

    Techniques: Isolation, Incubation, Western Blot

    Effects of garcinol and p300 siRNA on leptin-stimulated A549 cells. Serum-starved A549 cells were pretreated with the OB-R antibody (2 µg/mL), U0126 (10 µM), SB202190 (10 µM), SP600125 (10 µM), Bay11-7082 (10 µM), or garcinol (1 µM) for 1 h. Other cells were transfected with 100 nM p300 siRNA or scrambled siRNA as described in the Experimental section. The cells were then incubated with 1 µg/mL of leptin for the indicated time intervals. At the end of the incubation, the cells were harvested and cell lysates were extracted. ( A , B ) Expression of cPLA 2 -α and p300; ( C ) phosphorylation of p65; and ( D ) acetylation of histone H4 were detected using Western blot with specific antibodies. Cell membranes were stripped and reprobed with the anti-GAPDH antibody as internal controls. The data are expressed as mean ± SEM of 5 independent experiments ( n = 5). p

    Journal: International Journal of Molecular Sciences

    Article Title: Leptin Promotes cPLA2 Gene Expression through Activation of the MAPK/NF-κB/p300 Cascade

    doi: 10.3390/ijms161126045

    Figure Lengend Snippet: Effects of garcinol and p300 siRNA on leptin-stimulated A549 cells. Serum-starved A549 cells were pretreated with the OB-R antibody (2 µg/mL), U0126 (10 µM), SB202190 (10 µM), SP600125 (10 µM), Bay11-7082 (10 µM), or garcinol (1 µM) for 1 h. Other cells were transfected with 100 nM p300 siRNA or scrambled siRNA as described in the Experimental section. The cells were then incubated with 1 µg/mL of leptin for the indicated time intervals. At the end of the incubation, the cells were harvested and cell lysates were extracted. ( A , B ) Expression of cPLA 2 -α and p300; ( C ) phosphorylation of p65; and ( D ) acetylation of histone H4 were detected using Western blot with specific antibodies. Cell membranes were stripped and reprobed with the anti-GAPDH antibody as internal controls. The data are expressed as mean ± SEM of 5 independent experiments ( n = 5). p

    Article Snippet: PD98059, U0126, SB202190, SP600125, and garcinol were obtained from Biomol (Plymouth Meeting, PA, USA).

    Techniques: Transfection, Incubation, Expressing, Western Blot

    Effects of Bay11-7082 on leptin-regulated cPLA 2 -α expression and p65 phosphorylation. Serum-starved A549 cells were pretreated with different concentrations of Bay11-7082, OB-R (2 µg/mL), U0126 (10 µM), SB202190 (10 µM), SP600125 (10 µM), or Bay11-7082 (10 µM) for 1 h. The cells were then stimulated by 1 µg/mL of leptin for the indicated time intervals. At the end of incubation, the cells were harvested and cell lysates were extracted. ( A ) Expression of cPLA 2 -α and ( B ) Ser276 phosphorylation of p65 were detected using Western blot with specific antibodies. The cell membranes were stripped and reprobed with the anti-GAPDH antibody as internal controls. The data are expressed as mean ± SEM of five independent experiments ( n = 5). p

    Journal: International Journal of Molecular Sciences

    Article Title: Leptin Promotes cPLA2 Gene Expression through Activation of the MAPK/NF-κB/p300 Cascade

    doi: 10.3390/ijms161126045

    Figure Lengend Snippet: Effects of Bay11-7082 on leptin-regulated cPLA 2 -α expression and p65 phosphorylation. Serum-starved A549 cells were pretreated with different concentrations of Bay11-7082, OB-R (2 µg/mL), U0126 (10 µM), SB202190 (10 µM), SP600125 (10 µM), or Bay11-7082 (10 µM) for 1 h. The cells were then stimulated by 1 µg/mL of leptin for the indicated time intervals. At the end of incubation, the cells were harvested and cell lysates were extracted. ( A ) Expression of cPLA 2 -α and ( B ) Ser276 phosphorylation of p65 were detected using Western blot with specific antibodies. The cell membranes were stripped and reprobed with the anti-GAPDH antibody as internal controls. The data are expressed as mean ± SEM of five independent experiments ( n = 5). p

    Article Snippet: PD98059, U0126, SB202190, SP600125, and garcinol were obtained from Biomol (Plymouth Meeting, PA, USA).

    Techniques: Expressing, Incubation, Western Blot

    Effects of various inhibitors on leptin-stimulated cPLA 2 -α mRNA expression. Serum-starved A549 cells were pretreated with OB-R (2 µg/mL), PD98059 (10 µM), U0126 (10 µM), SB202190 (10 µM), SP600125 (10 µM), Bay11-7082 (10 µM), or garcinol (1 µM) for 1 h. The cells were then incubated with 1 µg/mL of leptin for 6 h. At the end of incubation, mRNA were extracted and used as templates of cDNA. Expression of cPLA 2 -α mRNA was detected using RT-PCR. Data are expressed as mean ± SEM of five independent experiments ( n = 5). p

    Journal: International Journal of Molecular Sciences

    Article Title: Leptin Promotes cPLA2 Gene Expression through Activation of the MAPK/NF-κB/p300 Cascade

    doi: 10.3390/ijms161126045

    Figure Lengend Snippet: Effects of various inhibitors on leptin-stimulated cPLA 2 -α mRNA expression. Serum-starved A549 cells were pretreated with OB-R (2 µg/mL), PD98059 (10 µM), U0126 (10 µM), SB202190 (10 µM), SP600125 (10 µM), Bay11-7082 (10 µM), or garcinol (1 µM) for 1 h. The cells were then incubated with 1 µg/mL of leptin for 6 h. At the end of incubation, mRNA were extracted and used as templates of cDNA. Expression of cPLA 2 -α mRNA was detected using RT-PCR. Data are expressed as mean ± SEM of five independent experiments ( n = 5). p

    Article Snippet: PD98059, U0126, SB202190, SP600125, and garcinol were obtained from Biomol (Plymouth Meeting, PA, USA).

    Techniques: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction

    SB202190 attenuation of cPLA 2 -α expression and p38 MAPK phosphorylation. Serum-starved A549 cells were pretreated with different concentrations of SB202190 or the OB-R antibody for 1 h and then stimulated by 1 µg/mL of leptin for the indicated time intervals. At the end of incubation, the cells were harvested and cell lysates were extracted. ( A ) Expression of cPLA 2 -α and ( B , C ) phosphorylation of p38 MAPK were detected using Western blot with specific antibodies. Cell membranes were stripped and reprobed with the anti-GAPDH antibody as internal controls. The data are expressed as mean ± SEM of five independent experiments ( n = 5). p

    Journal: International Journal of Molecular Sciences

    Article Title: Leptin Promotes cPLA2 Gene Expression through Activation of the MAPK/NF-κB/p300 Cascade

    doi: 10.3390/ijms161126045

    Figure Lengend Snippet: SB202190 attenuation of cPLA 2 -α expression and p38 MAPK phosphorylation. Serum-starved A549 cells were pretreated with different concentrations of SB202190 or the OB-R antibody for 1 h and then stimulated by 1 µg/mL of leptin for the indicated time intervals. At the end of incubation, the cells were harvested and cell lysates were extracted. ( A ) Expression of cPLA 2 -α and ( B , C ) phosphorylation of p38 MAPK were detected using Western blot with specific antibodies. Cell membranes were stripped and reprobed with the anti-GAPDH antibody as internal controls. The data are expressed as mean ± SEM of five independent experiments ( n = 5). p

    Article Snippet: PD98059, U0126, SB202190, SP600125, and garcinol were obtained from Biomol (Plymouth Meeting, PA, USA).

    Techniques: Expressing, Incubation, Western Blot

    TLR2-MAPK-ERK1/2 signaling cascade is required for the induction of VEGFA and Snail-1 by PCN033 ( A ) Phosphorylation of ERK1/2 along with PCN033 infection. The β-actin was detected as the loading control. ( B ) Densitometrical analysis of the ERK1/2 activation in hBMEC 2 h post-infection, compared with that in uninfected cells. Data are calculated as the ratio of phospho-ERK1/2 to total ERK1/2. ( C ) Effects of the MAPK signaling inhibitors on the PCN033-induced upregulation of VEGFA. U0126 (selective inhibitor of ERK1/2) and SB202190 (selective inhibitor of p38) could significantly decrease the PCN033-induced upregulation of VEGFA, while SP600125 (specific inhibitor of JNK) could not. ( D ) Effects of the MAPK signaling inhibitors on the PCN033-induced upregulation of Snail-1. Selective ERK1/2 inhibitor U0126 could completely block the PCN033-induced upregulation of Snail-1. ( E – F ) Snail-1 knocking-down via shRNA in hBMEC did not affect the induction of VEGFA by PCN033, while blocking VEGFA pathway significantly decreased the upregulation of Snail-1. ( G – H ) Effects of the VEGFR inhibitors on PCN033-induced activation of ERK1/2 and the densitometric analysis. ( I – J ) TLR2 agonist Pam3CSK4 induced the activation of ERK1/2 in a dose-dependent manner. ( K – L ) Pam3CSK4 dose-dependently induced the upregulation of Snail-1 and VEGFA in hBMEC. Results are expressed as mean ± SD from three independent assays.

    Journal: Oncotarget

    Article Title: Induction of VEGFA and Snail-1 by meningitic Escherichia coli mediates disruption of the blood-brain barrier

    doi: 10.18632/oncotarget.11696

    Figure Lengend Snippet: TLR2-MAPK-ERK1/2 signaling cascade is required for the induction of VEGFA and Snail-1 by PCN033 ( A ) Phosphorylation of ERK1/2 along with PCN033 infection. The β-actin was detected as the loading control. ( B ) Densitometrical analysis of the ERK1/2 activation in hBMEC 2 h post-infection, compared with that in uninfected cells. Data are calculated as the ratio of phospho-ERK1/2 to total ERK1/2. ( C ) Effects of the MAPK signaling inhibitors on the PCN033-induced upregulation of VEGFA. U0126 (selective inhibitor of ERK1/2) and SB202190 (selective inhibitor of p38) could significantly decrease the PCN033-induced upregulation of VEGFA, while SP600125 (specific inhibitor of JNK) could not. ( D ) Effects of the MAPK signaling inhibitors on the PCN033-induced upregulation of Snail-1. Selective ERK1/2 inhibitor U0126 could completely block the PCN033-induced upregulation of Snail-1. ( E – F ) Snail-1 knocking-down via shRNA in hBMEC did not affect the induction of VEGFA by PCN033, while blocking VEGFA pathway significantly decreased the upregulation of Snail-1. ( G – H ) Effects of the VEGFR inhibitors on PCN033-induced activation of ERK1/2 and the densitometric analysis. ( I – J ) TLR2 agonist Pam3CSK4 induced the activation of ERK1/2 in a dose-dependent manner. ( K – L ) Pam3CSK4 dose-dependently induced the upregulation of Snail-1 and VEGFA in hBMEC. Results are expressed as mean ± SD from three independent assays.

    Article Snippet: U0126, SB202190 and SP600125 were from Cayman Chemical Company (Ann Arbor, MI, USA).

    Techniques: Infection, Activation Assay, Blocking Assay, shRNA

    Suppression ­­­of p38 signaling restores sensitivity to PIM inhibition in primary AML cells and mouse xenografts A.-B. Inhibition of p38 enhances AZD1208 response in primary AML cells. Patient cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.313 μM, 1.25 μM, or 5 μM SCIO-469. Growth was measured after 4 days using MTT. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). C. Combined p38/PIM inhibition suppresses mTOR signaling. Primary AML cells were treated for 24 hours with 1 μM AZD1208, 10 μM SCIO-469, or the combination. Cell lysates were harvested and subjected to western blot analysis. D.-G. Inhibition of p38 enhances AZD1208 response in a validation set of primary AML cells. Cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.3 μM, 1 μM, or 3 μM SCIO-469. Viability was measured after 72 hours using CellTiter-Glo. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). H. SCIO-469 is synergistic with AZD1208. Primary AML cells were treated with 2-fold dilutions of AZD1208, SCIO-469, or the combination. Viability was assessed by CellTiter-Blue and used to calculate synergy scores. I. Dual PIM/p38 inhibition suppresses tumor growth in a xenograft model. K562 cells (5.10 6 ) were subcutaneously implanted in Rag2 −/− IL2γc −/− mice. Once tumors were established, animals were treated with vehicle, AZD1208 (30 mg/kg), SB202190 (5 mg/kg), or both drugs in combination. J. Mean tumor volume after 18 days treatment. Vehicle ( n = 11), AZD1208 ( n = 12), SB202190 ( n = 18), and combination ( n = 14). P -values were calculated via one-way ANOVA and Dunnett's test.

    Journal: Oncotarget

    Article Title: Intrinsic resistance to PIM kinase inhibition in AML through p38α-mediated feedback activation of mTOR signaling

    doi: 10.18632/oncotarget.9822

    Figure Lengend Snippet: Suppression ­­­of p38 signaling restores sensitivity to PIM inhibition in primary AML cells and mouse xenografts A.-B. Inhibition of p38 enhances AZD1208 response in primary AML cells. Patient cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.313 μM, 1.25 μM, or 5 μM SCIO-469. Growth was measured after 4 days using MTT. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). C. Combined p38/PIM inhibition suppresses mTOR signaling. Primary AML cells were treated for 24 hours with 1 μM AZD1208, 10 μM SCIO-469, or the combination. Cell lysates were harvested and subjected to western blot analysis. D.-G. Inhibition of p38 enhances AZD1208 response in a validation set of primary AML cells. Cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with 0.3 μM, 1 μM, or 3 μM SCIO-469. Viability was measured after 72 hours using CellTiter-Glo. A day 0 measurement was used as a baseline value (y-axis = ‘0’) for growth. Negative relative growth indicates a cytotoxic effect ( n = 2). H. SCIO-469 is synergistic with AZD1208. Primary AML cells were treated with 2-fold dilutions of AZD1208, SCIO-469, or the combination. Viability was assessed by CellTiter-Blue and used to calculate synergy scores. I. Dual PIM/p38 inhibition suppresses tumor growth in a xenograft model. K562 cells (5.10 6 ) were subcutaneously implanted in Rag2 −/− IL2γc −/− mice. Once tumors were established, animals were treated with vehicle, AZD1208 (30 mg/kg), SB202190 (5 mg/kg), or both drugs in combination. J. Mean tumor volume after 18 days treatment. Vehicle ( n = 11), AZD1208 ( n = 12), SB202190 ( n = 18), and combination ( n = 14). P -values were calculated via one-way ANOVA and Dunnett's test.

    Article Snippet: When tumor size reached ~50 to 100 mm3 , mice were randomly assigned and treated once daily with 30 mg/kg AZD1208 (AstraZeneca) by oral gavage and/or 5 mg/kg SB202190 (LC Laboratories) by intraperitoneal injection.

    Techniques: Inhibition, MTT Assay, Western Blot, Mouse Assay

    Pharmacological inhibition of p38 synergizes with AZD1208 through reduced mTOR signaling A.-E. p38 inhibitors enhance AZD1208 response. OCI-M1, OCI-M2, U2932, K562, and TMD8 cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with the p38 inhibitor SB202190. Viability was measured after 5 days using CellTiter-Blue ( n = 3). F. p38 inhibitors are synergistic with AZD1208. OCI-M1, OCI-M2, U2932, K562, and TMD8 cells were treated with 2-fold dilutions of AZD1208, SB202190, SCIO-469, or combinations for 5 days. Viability was assessed by CellTiter-Blue and used to calculate synergy scores. Self-self combination treatments were used as a baseline to determine significance ( n = 3). P-values were calculated using a one-way ANOVA and Dunnett's test. p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), and p ≤ 0.0001 (****) G. Combined inhibition of PIM and p38 results in reduced AKT/mTOR signaling after 48 hours. OCI-M1 (2.10 5 cells/well in 6-well plate) and OCI-M2 (4.10 5 cells/well in 6-well plate) cells were treated for 48 hours with 2 μM AZD1208, 20 μM SB202190, or the combination. Cell lysates were harvested and subjected to western blot analysis ( n = 3).

    Journal: Oncotarget

    Article Title: Intrinsic resistance to PIM kinase inhibition in AML through p38α-mediated feedback activation of mTOR signaling

    doi: 10.18632/oncotarget.9822

    Figure Lengend Snippet: Pharmacological inhibition of p38 synergizes with AZD1208 through reduced mTOR signaling A.-E. p38 inhibitors enhance AZD1208 response. OCI-M1, OCI-M2, U2932, K562, and TMD8 cells were treated with increasing concentrations of AZD1208 (x-axis) and co-treated with the p38 inhibitor SB202190. Viability was measured after 5 days using CellTiter-Blue ( n = 3). F. p38 inhibitors are synergistic with AZD1208. OCI-M1, OCI-M2, U2932, K562, and TMD8 cells were treated with 2-fold dilutions of AZD1208, SB202190, SCIO-469, or combinations for 5 days. Viability was assessed by CellTiter-Blue and used to calculate synergy scores. Self-self combination treatments were used as a baseline to determine significance ( n = 3). P-values were calculated using a one-way ANOVA and Dunnett's test. p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), and p ≤ 0.0001 (****) G. Combined inhibition of PIM and p38 results in reduced AKT/mTOR signaling after 48 hours. OCI-M1 (2.10 5 cells/well in 6-well plate) and OCI-M2 (4.10 5 cells/well in 6-well plate) cells were treated for 48 hours with 2 μM AZD1208, 20 μM SB202190, or the combination. Cell lysates were harvested and subjected to western blot analysis ( n = 3).

    Article Snippet: When tumor size reached ~50 to 100 mm3 , mice were randomly assigned and treated once daily with 30 mg/kg AZD1208 (AstraZeneca) by oral gavage and/or 5 mg/kg SB202190 (LC Laboratories) by intraperitoneal injection.

    Techniques: Inhibition, Western Blot

    Phospho-p38 MAPK inhibitors prevent and reverse EndoMT-associated gene expression in vitro. ( a ) To study possible prevention of EndoMT, hREC were treated with the p38 MAPK inhibitor SB203580 or SB202190 (10 μM) during EndoMT induction. Expression levels for the genes encoding the EndoMT-associated proteins SNAI1, SNAI2, α-SMA, FSP-1, vimentin, fibronectin, COL1A2 and COL3A1; and the endothelial differentiation markers VE-cadherin, CD31 and VEGFR2 were assessed by qPCR on day 6 of EndoMT induction. Data = mean ± SEM, * p

    Journal: Cells

    Article Title: Fibrotic Changes and Endothelial-to-Mesenchymal Transition Promoted by VEGFR2 Antagonism Alter the Therapeutic Effects of VEGFA Pathway Blockage in a Mouse Model of Choroidal Neovascularization

    doi: 10.3390/cells9092057

    Figure Lengend Snippet: Phospho-p38 MAPK inhibitors prevent and reverse EndoMT-associated gene expression in vitro. ( a ) To study possible prevention of EndoMT, hREC were treated with the p38 MAPK inhibitor SB203580 or SB202190 (10 μM) during EndoMT induction. Expression levels for the genes encoding the EndoMT-associated proteins SNAI1, SNAI2, α-SMA, FSP-1, vimentin, fibronectin, COL1A2 and COL3A1; and the endothelial differentiation markers VE-cadherin, CD31 and VEGFR2 were assessed by qPCR on day 6 of EndoMT induction. Data = mean ± SEM, * p

    Article Snippet: For the prevention study using the p38 MAPK inhibitors, SB203580 or SB202190 (10 μM, Abcam, Cambridge, MA, USA) was applied with the pro-inflammatory cytokines at the beginning of induction and again at day three; expression of endothelial and EndoMT markers was analyzed by RT-qPCR at day six.

    Techniques: Expressing, In Vitro, Real-time Polymerase Chain Reaction