p38mapk inhibitor sb202190  (STEMCELL Technologies Inc)

Journal: Pflugers Archiv : European journal of physiology

Article Title: Myostatin regulates the production of fibroblast growth factor 23 (FGF23) in UMR106 osteoblast-like cells.

doi: 10.1007/s00424-021-02561-y

Figure Lengend Snippet: Fig. 5 Schematic illustration of myostatin-induced FGF23 production in UMR106 cells. Myostatin binding to ACVR2B and following part- nering with TGF-βRI activates p38MAPK and NFκB. NFκB induces SOCE, resulting in induction of Fgf23 gene expression. Created with BioRender.com. Activin type 2 receptor B (ACVR2B); fibroblast growth factor 23 (FGF23); nuclear factor kappa-light-chain-enhancer of activated B-cells (NFκB); p38 mitogen–activated protein kinase (p38MAPK); store-operated Ca2 + entry (SOCE); transforming growth factor-β type I receptor (TGF-βRI)

Article Snippet: Next, cells were treated for further 24 h with recombinant myostatin protein (5–100 ng/mL, PeproTech, Rocky Hill, NJ, USA) in the presence or absence of TGF-β type I receptor (TGF-βRI) inhibitor SB431542 (10 μM, Sigma-Aldrich, Schnelldorf, Germany), p38MAPK inhibitor SB202190 (10 μM, Tocris), NFκB inhibitor withaferin A (500 nM, Tocris), or SOCE inhibitor 2-APB (150 μM, Sigma).

Techniques: Binding Assay, Gene Expression

Journal: Cell communication and signaling : CCS

Article Title: Platelet-derived microparticles stimulate the invasiveness of colorectal cancer cells via the p38MAPK-MMP-2/MMP-9 axis.

doi: 10.1186/s12964-023-01066-8

Figure Lengend Snippet: Fig. 5 Effect of PMPs on p38MAPK phosphorylation. A Representative Western blots of phosphorylated p38MAPK (phospho-p38) and total p38MAPK (p38) in CRC cells before and after 10 min of incubation with PMPs. B Normalized densitometric values of phospho-p38 and the total p38MAPK ratio. Quantified data are presented and statistically analysed as in Fig. 1. ***P < 0.001, *P < 0.05, N = 5

Article Snippet: The gelatinolytic properties of CRC cell were blocked by the appropriate inhibitors of MMP-2 (ARP101) or MMP-9 (CTK8G1150) at a final concentration of 20 μM or by the specific inhibitor of p38MAPK (SB202190, Cell Signaling Technology) at a final concentration of 5 μM.

Techniques: Phospho-proteomics, Western Blot, Incubation

Journal: Nephron Extra

Article Title: Crosstalk between Smad and Mitogen-Activated Protein Kinases for the Regulation of Apoptosis in Cyclosporine A- Induced Renal Tubular Injury

doi: 10.1159/000333014

Figure Lengend Snippet: Effect of inhibition of MAPKs in the presence or absence of EGF on CyA-induced nuclear translocation of R-Smad. CyA (42 μ M ) increased the expression of p-Smad2 (lane 2) at 6 h, measured by Western blotting, compared to control cells (C, lane 1). SB202190 (SB; 20 μ M ) deteriorated nuclear translocation of p-Smad2 in both control (lane 4) and CyA-treated cells (lane 5), whereas PD98059 (PD; 20 μ M ) ameliorated nuclear translocation of p-Smad2 in CyA-treated cells (lane 7). EGF (20 ng/ml) ameliorated CyA-induced nuclear translocation of p-Smad2 (lane 3), which was abolished by SB202190 (lane 6) but not by PD98059 (lane 8).

Article Snippet: The following reagents and antibodies were purchased: fetal bovine serum, Dulbecco's modified Eagle's medium/nutrient mix F12 (DMEM/F12) and Alexa Fluor® 488 goat anti-rabbit antibody (Invitrogen, Carlsbad, Calif., USA); p38MAPK inhibitor, SB202190, and Hoechst 33258 (Sigma, St. Louis, Mo., USA); antibodies for p-Smad2, p-Smad3, Smad2, Smad3, p-ERK1/2, ERK1/2, p-p38MAPK, p38MAPK, p-JNK1/2/3, histone H3, and an inhibitor of ERK, PD98059 (Cell Signaling Technology, Beverly, Mass., USA); recombinant human EGF (R&D Systems, Minneapolis, Minn., USA); Fluoromount (Diagnostic Biosystems, Pleasanton, Calif., USA); protein assay kit (Bio-Rad, Hercules, Calif., USA); Amersham ECL Western blotting detection reagents (GE Healthcare, Little Chalfont, UK), and Lab-Tek II 8-chamber glass slides (Nunc, Naperville, Ill., USA).

Techniques: Inhibition, Translocation Assay, Expressing, Western Blot

Journal: Nephron Extra

Article Title: Crosstalk between Smad and Mitogen-Activated Protein Kinases for the Regulation of Apoptosis in Cyclosporine A- Induced Renal Tubular Injury

doi: 10.1159/000333014

Figure Lengend Snippet: Effect of inhibition of MAPKs on CyA-induced apoptosis. PD98059 (PD; 20 μ M ) but not SB202190 (SB; 20 μ M ) ameliorated CyA (42 μ M )-induced apoptosis at 24 h. Data are expressed as means ± SEM. n = 3, * p < 0.02, ** p < 0.001.

Article Snippet: The following reagents and antibodies were purchased: fetal bovine serum, Dulbecco's modified Eagle's medium/nutrient mix F12 (DMEM/F12) and Alexa Fluor® 488 goat anti-rabbit antibody (Invitrogen, Carlsbad, Calif., USA); p38MAPK inhibitor, SB202190, and Hoechst 33258 (Sigma, St. Louis, Mo., USA); antibodies for p-Smad2, p-Smad3, Smad2, Smad3, p-ERK1/2, ERK1/2, p-p38MAPK, p38MAPK, p-JNK1/2/3, histone H3, and an inhibitor of ERK, PD98059 (Cell Signaling Technology, Beverly, Mass., USA); recombinant human EGF (R&D Systems, Minneapolis, Minn., USA); Fluoromount (Diagnostic Biosystems, Pleasanton, Calif., USA); protein assay kit (Bio-Rad, Hercules, Calif., USA); Amersham ECL Western blotting detection reagents (GE Healthcare, Little Chalfont, UK), and Lab-Tek II 8-chamber glass slides (Nunc, Naperville, Ill., USA).

Techniques: Inhibition

Journal: Nephron Extra

Article Title: Crosstalk between Smad and Mitogen-Activated Protein Kinases for the Regulation of Apoptosis in Cyclosporine A- Induced Renal Tubular Injury

doi: 10.1159/000333014

Figure Lengend Snippet: Immunofluorescence analysis for the effect of EGF on CyA-induced nuclear translocation of R-Smad and its modulation by inhibition of MAPKs. Immunofluorescence analysis showed that CyA induced nuclear translocation of p-Smad3 at 6 h ( d , arrows) compared to control cells ( a ). EGF did not affect nuclear translocation of p-Smad3 in control cells ( b ). EGF ameliorated CyA-induced translocation of p-Smad3 ( c ), which was abolished by SB202190 ( e , arrows) but not by PD98059 ( f ). Data are representative of 3 independent experiments.

Article Snippet: The following reagents and antibodies were purchased: fetal bovine serum, Dulbecco's modified Eagle's medium/nutrient mix F12 (DMEM/F12) and Alexa Fluor® 488 goat anti-rabbit antibody (Invitrogen, Carlsbad, Calif., USA); p38MAPK inhibitor, SB202190, and Hoechst 33258 (Sigma, St. Louis, Mo., USA); antibodies for p-Smad2, p-Smad3, Smad2, Smad3, p-ERK1/2, ERK1/2, p-p38MAPK, p38MAPK, p-JNK1/2/3, histone H3, and an inhibitor of ERK, PD98059 (Cell Signaling Technology, Beverly, Mass., USA); recombinant human EGF (R&D Systems, Minneapolis, Minn., USA); Fluoromount (Diagnostic Biosystems, Pleasanton, Calif., USA); protein assay kit (Bio-Rad, Hercules, Calif., USA); Amersham ECL Western blotting detection reagents (GE Healthcare, Little Chalfont, UK), and Lab-Tek II 8-chamber glass slides (Nunc, Naperville, Ill., USA).

Techniques: Immunofluorescence, Translocation Assay, Inhibition

Journal: Nephron Extra

Article Title: Crosstalk between Smad and Mitogen-Activated Protein Kinases for the Regulation of Apoptosis in Cyclosporine A- Induced Renal Tubular Injury

doi: 10.1159/000333014

Figure Lengend Snippet: Effect of EGF in the presence or absence of inhibition of MAPKs on CyA-induced apoptosis. EGF (20 ng/ml) rescued CyA (42 μ M )-induced apoptosis at 24 h. SB202190 (SB; 20 μ M ) but not PD98059 (PD; 20 μ M ) abolished the protective effect of EGF on CyA-induced apoptosis. Data are expressed as means ± SEM. n = 3, * p < 0.02, ** p < 0.001.

Article Snippet: The following reagents and antibodies were purchased: fetal bovine serum, Dulbecco's modified Eagle's medium/nutrient mix F12 (DMEM/F12) and Alexa Fluor® 488 goat anti-rabbit antibody (Invitrogen, Carlsbad, Calif., USA); p38MAPK inhibitor, SB202190, and Hoechst 33258 (Sigma, St. Louis, Mo., USA); antibodies for p-Smad2, p-Smad3, Smad2, Smad3, p-ERK1/2, ERK1/2, p-p38MAPK, p38MAPK, p-JNK1/2/3, histone H3, and an inhibitor of ERK, PD98059 (Cell Signaling Technology, Beverly, Mass., USA); recombinant human EGF (R&D Systems, Minneapolis, Minn., USA); Fluoromount (Diagnostic Biosystems, Pleasanton, Calif., USA); protein assay kit (Bio-Rad, Hercules, Calif., USA); Amersham ECL Western blotting detection reagents (GE Healthcare, Little Chalfont, UK), and Lab-Tek II 8-chamber glass slides (Nunc, Naperville, Ill., USA).

Techniques: Inhibition

Journal: Molecular Medicine Reports

Article Title: Hydrogen sulfide improves ox-LDL-induced expression levels of Lp-PLA 2 in THP-1 monocytes via the p38MAPK pathway

doi: 10.3892/mmr.2021.11997

Figure Lengend Snippet: H 2 S inhibits ox-LDL-induced expression levels of Lp-PLA 2 by blocking p38MAPK. Cells were pretreated with the specific p38 inhibitors SB203580 (20 mM) and SB202190 (20 mM) for 30 min, with PPG (3 mM) for 2 h or with NaHS (100 µM) for 24 h prior to incubation with ox-LDL (50 µg/ml) for 24 h, after which (A) protein expression levels of p-p38MAPK and t-p38MAPK, and relative (B) protein and (C) mRNA Lp-PLA 2 levels were determined via western blot analysis. (D) Lp-PLA 2 activity was determined using a Lp-PLA 2 activity kit. Data are presented as the mean ± SEM (n=5). ! P<0.01 vs. control; *P<0.05 vs. ox-LDL; **P<0.01 vs. ox-LDL + PPG. ox-LDL, oxidized low-density lipoprotein; Lp-PLA 2 , lipoprotein-associated phospholipase A2; PPG, DL-propargylglycine; t-, total; p-, phosphorylated.

Article Snippet: RPMI-1640 cell culture medium (cat. no. 72400120; Thermo Fisher Scientific, Inc.) and FBS were obtained from Gibco (cat. no.12483020; Thermo Fisher Scientific, Inc.). ox-LDL, the p38MAPK specific inhibitors SB203580 (cat. no. S8307) and SB202190 (cat. no. S7067), NaHS (cat. no. 161527) and DL-propargylglycine (PPG; cat. no. P7888) were purchased from Sigma-Aldrich (Merck KGaA).

Techniques: Expressing, Blocking Assay, Incubation, Western Blot, Activity Assay

Journal: Molecular Medicine Reports

Article Title: Hydrogen sulfide improves ox-LDL-induced expression levels of Lp-PLA 2 in THP-1 monocytes via the p38MAPK pathway

doi: 10.3892/mmr.2021.11997

Figure Lengend Snippet: H 2 S decreases lipid accumulation in macrophages by inhibiting the activity of Lp-PLA 2 . THP-1 cells were preincubated with PMA for 72 h to establish a macrophage model. THP-1 macrophages were pretreated with SB203580 (20 mM) and SB202190 (20 mM) for 30 min, PPG (3 mM) for 2 h, NaHS (100 µM) for 24 h or Lp-PLA 2 siRNA (30 nM) for 48 h, prior to incubation with ox-LDL (50 µg/ml) for 24 h. Cells exhibiting lipid accumulation were observed and counted via light microscopy following ORO staining (magnification, ×40) in the (A) Control, (B) ox-LDL, (C) ox-LDL + PPG, (D) ox-LDL + SB203580, (E) ox-LDL + SB202190, (F) ox-LDL + NaHS and (G) ox-LDL + Lp-PLA 2 siRNA groups. Images are representative of six independent repeats. (H) Quantitative analysis of the positive area of ORO (%) in each group. Detection of Lp-PLA 2 siRNA transfection by (I) western blotting and (J) RT-PCR. THP-1 macrophages were pretreated with Lp-PLA 2 siRNA (30 nM) or NC siRNA(30 nM) for 48 h. (K) Western blotting and (L) RT-PCR were conducted to detect the effect of Lp-PLA 2 siRNA on inhibiting the expression of Lp-PLA 2 in THP-1 macrophages induced by ox-LDL. ! P<0.01 vs. control; *P<0.05 vs. ox-LDL; **P<0.01 vs. ox-LDL + PPG; & P<0.01 vs. ox-LDL + Lp-PLA 2 siRNA. Lp-PLA 2 , lipoprotein-associated phospholipase A2; PMA, phorbol-12-myristate-13-acetate; PPG, DL-propargylglycine; ox-LDL, oxidized low-density lipoprotein; ORO, oil red O; RT, reverse transcription; siRNA, small interfering RNA; NC, negative control.

Article Snippet: RPMI-1640 cell culture medium (cat. no. 72400120; Thermo Fisher Scientific, Inc.) and FBS were obtained from Gibco (cat. no.12483020; Thermo Fisher Scientific, Inc.). ox-LDL, the p38MAPK specific inhibitors SB203580 (cat. no. S8307) and SB202190 (cat. no. S7067), NaHS (cat. no. 161527) and DL-propargylglycine (PPG; cat. no. P7888) were purchased from Sigma-Aldrich (Merck KGaA).

Techniques: Activity Assay, Incubation, Light Microscopy, Staining, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Small Interfering RNA, Negative Control