Fig. 4

GC cell-derived activin B regulates NF-κB activity of fibroblasts through ALK7/TRAF6/TAK1. A-B. KEGG pathway enrichment analysis of significantly differential expression genes (DEGs). C. Co-culture with activin B, stable knockdown, or overexpression of INHBB in GC cells affected protein levels of TRAF6 and phosphorylation level TAK1 in normal gastric fibroblasts. D. Normal gastric fibroblasts stimulated by activin B or treated by control reagent or SB-431,542. Cell lysates were immunoprecipitated with an anti-TRAF6 antibody and were followed by immunoblotting with anti-ubiquitin antibody. E. Western blot analyses of the levels of p-IKKα/β, p-IκBα, p-p65, total IKKα/β, IκBα and p65 in fibroblast co-cultured with activin B or different INHBB-expressing GC cells. F. Immunofluorescence assay evaluating the nuclear translocation of p65 in fibroblast co-cultured with activin B or different INHBB expressing GC cells. G. Immunofluorescence assay evaluating the effect of JSH-23 to the nuclear translocation of p65 in fibroblasts. H. Fibroblasts under high activin B environment were treated with JSH-23 as indicated, and cell proliferation was verified by CCK-8 assay (n = 3). I. Fibroblasts under high activin B environment were treated with JSH-23. Transwell assay was used to evaluate cell migration and invasion (n = 3). **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. rActivin B: Recombinant Human Activin B; Sup.: supernatant