Fig. 9

Effect of MLL3 shRNA on synaptopodin and cathepsin L expression and histone H3K4 me3 levels and the determination of H3K4 me3 level on the promoter region of cathepsin L in cultured mouse podocytes co-cultured with LPS-stimulated peritoneal macrophages. In podocytes co-cultured with peritoneal macrophages stimulated with LPS, increased H3K4 me3 was found compared to untreated podocytes (Veh) (a). MLL3 shRNA administration decreased H3K4 me3 compared to that of cells administered with control shRNA (a). MLL3 shRNA administration also decreased the level of cathepsin L protein expression compared to that of cells administered the control shRNA (b). MLL3 shRNA increased the expression of synaptopodin compared to that of the cells administered control shRNA in cultures stimulated with cytokines derived from co-cultured macrophages (c). In chromatin immunoprecipitation (ChIP) assays with E11 cells stimulated with cytokines derived from co-cultured macrophages, the stimulated levels of H3K4 me3 at the cathepsin L promoters were significantly lower in E11 cells treated with MLL3 shRNA 72 h post-administration compared with the control shRNA (d). *P < 0.05 vs. Veh. Veh, untreated podocytes; Cont, control shRNA; H3K4 me3, histone H3K4 trimethylation; total H3, total histone H3; N = 4. Data are expressed as the mean ± SE