p38 mapk pathway inhibitor sb203580  (MedChemExpress)

Journal: American Journal of Translational Research

Article Title: Morinda officinalis polysaccharide enhances osteogenic differentiation and migration of bone marrow mesenchymal cells by activating P38MAPK signal transduction

doi: 10.62347/VVPN2529

Figure Lengend Snippet: Effects of MOP on MAPK signaling pathway. A. Phosphorylation of p38, ERK1/2, and JNK in BMSCs analyzed using western blotting. B. Quantitative results of ERK1/2 phosphorylation. C. Quantitative results of p38 phosphorylation. D. Quantitative results of JNK phosphorylation. * P <0.05, ** P <0.01.

Article Snippet: Once the monolayer of cells covered 90% of the bottom of the wells, the medium was replaced with fresh medium containing MOP (Shifengbio, Shanghai, China) or MOP combined with the p38 MAPK pathway inhibitor SB203580 (2 μg/mL, HY-10256, MedChemExpress, Monmouth Junction, NJ, USA), and incubated for 24 h [ 19 ].

Techniques: Phospho-proteomics, Western Blot

Journal: American Journal of Translational Research

Article Title: Morinda officinalis polysaccharide enhances osteogenic differentiation and migration of bone marrow mesenchymal cells by activating P38MAPK signal transduction

doi: 10.62347/VVPN2529

Figure Lengend Snippet: Effects of P38 MAPK signaling pathway inhibitor and MOP on MSC proliferation. A. BMSC viability evaluated using MTT assay. B, C. BMSC clone formation rate evaluated using colony formation experiment (10×, 200 μm). D, E. BMSC migration assessed using wound-healing assay. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: Once the monolayer of cells covered 90% of the bottom of the wells, the medium was replaced with fresh medium containing MOP (Shifengbio, Shanghai, China) or MOP combined with the p38 MAPK pathway inhibitor SB203580 (2 μg/mL, HY-10256, MedChemExpress, Monmouth Junction, NJ, USA), and incubated for 24 h [ 19 ].

Techniques: MTT Assay, Migration, Wound Healing Assay

Journal: American Journal of Translational Research

Article Title: Morinda officinalis polysaccharide enhances osteogenic differentiation and migration of bone marrow mesenchymal cells by activating P38MAPK signal transduction

doi: 10.62347/VVPN2529

Figure Lengend Snippet: Effect of P38 MAPK signaling pathway inhibitor and MOP on BMSC osteogenic differentiation. A, B. Alizarin red staining for detection of calcium deposits (20×, 100 μm). C, D. Alkaline phosphatase (ALP) staining for detection of enzyme activity (20×, 100 μm). E-I. The protein expression levels of collagen I (Col I), osteocalcin (OCN), runt-related transcription factor 2 (RUNX2) and ALP levels examined using western blot. J-M. The mRNA expression levels of OCN, ALP, RUNX2, and Col I in BMSCs assessed using RT-qPCR. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: Once the monolayer of cells covered 90% of the bottom of the wells, the medium was replaced with fresh medium containing MOP (Shifengbio, Shanghai, China) or MOP combined with the p38 MAPK pathway inhibitor SB203580 (2 μg/mL, HY-10256, MedChemExpress, Monmouth Junction, NJ, USA), and incubated for 24 h [ 19 ].

Techniques: Staining, Activity Assay, Expressing, Western Blot, Quantitative RT-PCR

Journal: Advanced Biology

Article Title: HMGB1 Derived from the Pyroptotic Microenvironment Promotes Macrophage Extracellular Traps in Hirschsprung‐Associated Enterocolitis

doi: 10.1002/adbi.202400761

Figure Lengend Snippet: The TLR4‐P38 MAPK/P65 NF‐kB signaling pathways mediate the‐pyroptotic‐microenvironment‐induced MET formation. A) Western blot analysis of p‐ERK, p‐p38, p‐JNK and p‐p65 levels in macrophages cocultured with pyroptotic‐CM for 4 h. B,C) Macrophages were pretreated with inhibitors of the ERK, JNK, p38 MAPK, and p65 NF‐kB pathways prior to incubation with pyroptotic‐CM. The MET formation by macrophages was evaluated with SYTOX Green staining and detected by flow cytometry. The representative images are shown in B and the quantification of B is shown in C (one‐way ANOVA followed by Tukey's multiple‐comparison test, n = 3 per group). D) Macrophages were pretreated with inhibitors of TLR2, TLR4, TLR9 and RAGE prior to coculture with pyroptotic‐CM, and the p‐p38 and p‐p65 levels in macrophages were measured by western blotting. E,F) The MET formation of macrophages pretreated with inhibitors of TLR2, TLR4 and RAGE prior to incubation with pyroptotic‐CM was evaluated with SYTOX Green staining and detected by flow cytometry. The representative images are shown in E and the quantification of E is shown in F (one‐way ANOVA followed by Tukey's multiple‐comparison test, n = 3 per group). Data are expressed as mean ± SD. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The inhibitors used in this study included an HMGB1 antagonist (HY‐N0184, MCE, USA), a p38 MAPK pathway inhibitor (HY‐12839, MCE, USA), a p65 NF‐kB pathway inhibitor (HY‐138537, MCE, USA), an ERK pathway inhibitor (HY‐112287, MCE, USA), a JNK inhibitor (HY‐12041, MCE, USA), a TLR2 antagonist (HY‐112146, MCE, USA), a TLR4 antagonist (HY‐11109, MCE, USA), a TLR9 antagonist ( HY131952 , MCE, USA), and a RAGE antagonist (HY‐P2268).

Techniques: Protein-Protein interactions, Western Blot, Incubation, Staining, Flow Cytometry, Comparison

Journal: Advanced Biology

Article Title: HMGB1 Derived from the Pyroptotic Microenvironment Promotes Macrophage Extracellular Traps in Hirschsprung‐Associated Enterocolitis

doi: 10.1002/adbi.202400761

Figure Lengend Snippet: HMGB1 induces MET formation through TLR4‐P38 MAPK/P65 NF‐kB signaling pathways in macrophages. A‐C) BMDMs isolated from mice were stimulated with HMGB1 or PBS. Then, differentially expressed genes (DEGs) were analyzed by RNA sequencing. (A) The number of DEGs in the HMGB1 group vs. the PBS group. Red represented upregulated DEGs, and blue downregulated DEGs. (B) KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analyses of the upregulated DEGs. The dot size represents the number of DEGs, and the dot color represents the corresponding p value. (C) Scatter plot showing DEGs in the HMGB1 group vs. the PBS group. Genes were plotted based on their expression levels. Red and green dots represented up and downregulated genes, respectively. D‐I) qRT‐PCR analysis of the indicated genes in macrophages treated with HMGB1 or PBS (Unpaired t‐test, n = 3 per group). J) Western blot analysis of p‐ERK, p‐p38, p‐JNK and p‐p65 levels in macrophages cocultured with HMGB1. K) Macrophages were pretreated with inhibitors of TLR4 prior to incubation with HMGB1, and the p‐p38 and p‐p65 levels in macrophages was measured by western blotting. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: The inhibitors used in this study included an HMGB1 antagonist (HY‐N0184, MCE, USA), a p38 MAPK pathway inhibitor (HY‐12839, MCE, USA), a p65 NF‐kB pathway inhibitor (HY‐138537, MCE, USA), an ERK pathway inhibitor (HY‐112287, MCE, USA), a JNK inhibitor (HY‐12041, MCE, USA), a TLR2 antagonist (HY‐112146, MCE, USA), a TLR4 antagonist (HY‐11109, MCE, USA), a TLR9 antagonist ( HY131952 , MCE, USA), and a RAGE antagonist (HY‐P2268).

Techniques: Protein-Protein interactions, Isolation, RNA Sequencing, Expressing, Quantitative RT-PCR, Western Blot, Incubation

Journal: Immunobiology

Article Title: Giardia duodenalis triggered neutrophil extracellular traps in goats.

doi: 10.1016/j.imbio.2025.152894

Figure Lengend Snippet: Fig. 6. NETs release and ROS production was associated with NADPH oxidase, ERK1/2 and p38 MAPK signaling pathway (A) NETs quantitation. (B) ROS level. Goat neutrophils were pretreated with NADPH oxidase inhibitor DPI (10 μM), ERK1/2 inhibitor U0126 (50 μM), p38 MAPK signaling pathway inhibitor SB202190 (10 μM) respectively for 30 min, and stimulated with Giardia (ratio 1:1) for 2 h. Zymosan (1 mg/mL) was used as a positive control. Data were expressed as mean ± SD (n = 5). P < 0.05 was considered significant (*P < 0.05, ***P < 0.001).

Article Snippet: In inhibition tests, prior to stimulation with G. duodenalis (at a ratio of 1:1), neutrophils were pretreated with 10 μM of the NADPH oxidase inhibitor Diphenyleneiodonium chloride (DPI, Sigma-Aldrich, USA), 50 μM of ERK1/2 inhibitor U0126 (Sigma-Aldrich, USA), 10 μM of the p38 MAPK signaling pathway inhibitor SB202190 (Sigma-Aldrich, USA), 100 μM of TLR2 inhibitor C29 (MedChemExpress, USA) and 10 μM of TLR4 inhibitor TLR4-IN-C34 (C34, MedChemExpress, USA) respectively for 30 min 30 min. Zymosan was used as the positive control at a concentration of 1 mg/mL.

Techniques: Quantitation Assay, Positive Control