Fig. 4

miR-205-5p elevates IGFBP3 expression by regulating the methylation of H3 histones by binding to EHMT2. A, B The expression of IGFBP3 in clinical tissues or PA-treated fibrotic cells using RT-qPCR and western blot analysis. C Prediction analysis of the UCSC database. D, E ChIP assay was implemented to detect the H3K9me2 and H3K4me3 enrichment in IGFBP3 promoter region in clinical tissues and cell models. F, G. EHMT2 and IGFBP3 expression levels in cells transfected with sh-EHMT2 were tested by RT-qPCR and western blot analysis. H, I H3K9me2 enrichment and H3K4me3 enrichment in IGFBP3 promoter region in cells transfected with sh-EHMT2 were assessed by ChIP assay. J, K Expression of IGFBP3 in cells co-transfected with miR-205-5p-mimic and sh-IGFBP3 was determined using RT-qPCR and western blot analysis. L TGF-β1 expression in cells co-transfected with miR-205-5p-mimic and sh-IGFBP3 was determined by RT-qPCR. M The secretion of cell inflammatory factors TNF-α and IL-1β in cells co-transfected with miR-205-5p-mimic and sh-IGFBP3 was measured by ELISA. N Expression of fibrosis-related proteins α-SMA, Col I and COL III in cells co-transfected with miR-205-5p-mimic and sh-IGFBP3 was determined by western blot analysis. *p < 0.05, vs. the SR group, the control group, the sh-NC group or the miR-205-5p-mimic + sh-NC group