Fig. 6

H19 facilitated peritoneal fibrosis by binding p300 and acetylating H3K27 in the VEGFA promoter region. a RNA FISH showed the localization of H19 in MeT-5A cells. An 18 s probe was used in the positive control. Scale bars = 100 μm. b Quantification of H19 in the cytoplasm and nucleus by cytoplasmic and nuclear RNA isolation experiments. c qRT-PCR measured the VEGFA mRNA levels in MeT-5A cells transfected with siH19 or siNC and stimulated with HG. n = 3. **P < 0.01 vs. siNC group, ^##P < 0.01 vs. siNC + HG group. d qRT-PCR to test LV-NC- or LV-H19-infected MeT-5A cells. n = 3. **P < 0.01 vs. LV-NC group. e–f ChIP assay to detect H3K27ac and p300 binding to the VEGFA promoter region in H19 knockdown MeT-5A cells. n = 3. **P < 0.01 vs. siNC group, ^##P < 0.01 vs. siNC + HG group. g and h ChIP assay to detect H3K27ac and p300 binding to the VEGFA promoter region in LV-H19 overexpression MeT-5A cells. n = 3. **P < 0.01 vs. LV-NC group. i and j ChIP assays were performed to detect H3K27ac and p300 binding to the VEGFA promoter region in MeT-5A cells under HG stimulation and TAM treatment. n = 3. **P < 0.01 vs. CON group, ^##P < 0.01 vs. HG group. k and l RIP assay showing p300-bound H19 in MeT-5A cells and mouse peritoneal primary cells. RNA enrichment was determined by qRT-PCR and normalized to the input control. IgG and Actin RNA were used as negative controls. n = 3. **P < 0.01 vs. H19/p300 group. There is no data significance between Actin/p300 and Actin/IgG group. Two-way ANOVA was performed followed by Dunnett's T3. m and n RNA pull-down was used to analyze protein binding to H19. RNA-binding proteins were enriched and analyzed by Western blotting. All experiments were performed three times. Values are the mean ± SD. Two-way ANOVA was performed followed by Tukey's test (e-i)