EP300 drives renal fibrosis in diabetic nephropathy via histone acetyltransferase-mediated nephrocystin-4 expression

Figure 2 EP300 regulates histone acetylation levels in nephrocystin-4 chromatin fragments. A: EP300 mRNA expression was measured using real-time quantitative polymerase chain reaction (RT-qPCR); B: Venny diagram of the intersection of EP300-regulated histone acetylation modification action targets with renal fibrosis-related targets; C: Chromatin immunoprecipitation (ChIP)-seq analysis to identify the key target nephrocystin-4 (NPHP4); D: NPHP4 gene abundance in ChIP products was measured using RT-qPCR; E: Measurement of EP300 interference efficiency using RT-qPCR; F and G: Interference efficiency of EP300 was measured using western blot; H and I: Representative western blot images and quantitative analysis of histone H3 lysine 27 protein levels; J: Enrichment of histone H3 lysine 27 at the NPHP4 locus was measured by ChIP-RT-qPCR in HK-2 cells transfected with siCtrl or small interfering RNA targeting EP300 under high glucose conditions. Data are presented as mean ± SD (n = 3). ^aP < 0.05, ^bP < 0.01, and ^cP < 0.001. ChIP: Chromatin immunoprecipitation; DN: Diabetic nephropathy; HG: High glucose; H3K27ac: Acetylated histone H3 lysine 27; IgG: Immunoglobulin G; siEP300: Small interfering RNA targeting EP300.