Sinefungin ameliorates the TGF-β1-induced increase of α-SMA and inhibits the upregulation of histone H3K4 mono-methylation in renal epithelial cells. Pretreatment of sinefungin (0.5 or 1.0 μg/mL) was conducted 60 minutes before TGF-β1 (10 ng/mL) stimulation. (A) A typical western blot for α-SMA in NRK-52E cells. Quantification is shown in the lower panel (n=5 for each group). (B) Representative western blot analysis of the expression of histone H3K4me1, (C) H3K4me2, and (D) H3K4me3 expression in NRK-52E cells. Quantification is shown in the lower panel (n=5 for each group). (E) Representative ChIP assay analysis of the expression of binding of H3K4me1 protein to Col1a1, CTGF, and PAI-1 promoters in NRK-52E cells. ChIP assays were performed with H3K4me1 antibody. Immunoprecipitated DNA and input DNA were subjected to qRT-PCR. Results were normalized to input DNA (n=5 for each group). Data were analyzed by one-way ANOVA followed by the post hoc test using t test with Bonferroni correction. #P<0.05; ##P<0.01. Sine, sinefungin.