Fig. 5

Activated fibroblasts regulate GC cell function via the IL-1β/p65 pathway. (A) A Venn diagram summarized the up-regulated cytokine genes in fibroblasts stimulated by exogenous activin B or co-cultured with GC cells (HGC-27 shNC vs. HGC-27 shINHBB; MGC-803 vs. MGC-803 shINHBB; MKN-45 INHBB vs. MKN-45 Vector). (B) mRNA levels of IL-1b was examined by qRT-PCR in GC cells (HGC-27 shNC/HGC-27 shINHBB, MGC-803/MGC-803 shINHBB, MKN-45 Vector/MKN-45 INHBB, HGC-27 Vector/HGC-27 INHBB). (C) Western blot analyses of the levels of p-p65 and p-65 in GC cells co-cultured with fibroblasts. (D) Western blot analyses of the levels of p-p65 and p-65 in GC cells treated with IL-1b for different times. (E) Cell proliferation was verified by CCK-8 assay. The value of the absorbance (at 450 nm) was recorded from 0 to 96 h in GC cells treated or untreated with IL-1b (n = 3). (F) Cell migration and invasion were detected by transwell assay in GC cells with or without IL-1b co-culture (n = 3). (G) Western blot analyses of the levels of p-p65 and p-65 in GC cells treated and untreated with JSH-23 in fibroblasts co-cultured system. (H) Cell proliferation was verified by CCK-8 assay. The value of the absorbance (at 450 nm) was recorded from 0 to 96 h in GC cells treated or untreated with JSH-23 in fibroblasts co-cultured system (n = 3). (I) Cell migration and invasion were detected by transwell assay in GC cells in fibroblasts co-cultured system with or without JSH-23 co-culture (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. rActivin B: Recombinant Human Activin B; Sup.: supernatant