Fig. 4.

DOT1L regulates the expression and activation of EGFR in peritoneal mesothelial cells. a HPMCs were transfected with scrambled siRNA and DOT1L siRNA for 6 h, and then incubated with 2ng/ml TGF-β1 for an additional 36 h before being harvested for analysis. Cell lysates were subjected to immunoblotting analysis with antibodies against PDGFRα, IGFRβ, FGFR1, Src, EGFR and GAPDH. b HPMCs were transfected with Flag-DOT1L pcDNA3.0 plasmid or vector for 0, 18, 36 h and then subjected to immunoblotting analysis with antibodies against DOT1L, H3K79me2, Histone H3, and GAPDH c HPMCs were transfected with Flag-DOT1L pcDNA3.0 plasmid or vector for 0, 18, 36 h and then subjected to immunoblotting analysis with antibodies against PDGFRα, IGFRβ, FGFR1, Src, EGFR, p-EGFR and GAPDH. d ChIP-qPCR analysis of H3K79me2 enrichment in the EGFR promoter region. e 2ng/ml TGF-β1-treated HPMCs were subjected to immunoprecipitation with IgG or DOT1L antibody, followed by EGFR and DOT1L immunoblotting. Input lysates were analyzed by EGFR, DOT1L and Histone H3 immunoblotting. f Immunofluorescence co-staining of DOT1L and EGFR in TGF-β1-treated HPMCs. Scale bar = 50μm. Data are means ± sem of 3 samples, *P < 0.05, **P < 0.01, *** P < 0.001, P ≥ 0.05 is not significant (NS)