p38 mapk signaling pathway  (MedChemExpress)

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