Fig. 5.

DOT1L inhibitor EPZ5676 reduces the expression and activation of EGFR in peritoneal mesothelial cells. a Serum-starved HPMCs were cultured in 2ng/ml TGF-β1 for 36 h with different concentrations of EPZ5676 (0, 1, 5, 10 μM). Cell lysates were subjected to immunoblotting analysis with specific antibodies against DOT1L, H3K79me2, Histone H3 or GAPDH. b Expression levels of DOT1L and H3K79me2 were quantified by densitometry and normalized with GAPDH, Histone H3, respectively. c Cell lysates were subjected to immunoblotting analysis with specific antibodies against p-EGFR, EGFR or GAPDH. d Expression levels of p-EGFR and EGFR were quantified by densitometry and normalized with EGFR and GAPDH, respectively. e Immunofluorescence of EGFR in HPMCs under different treatments (starved, 2ng/ml TGF-β1 or 60mM HG) with or without 10μM EPZ5676. f Immunofluorescence of p-EGFR in HPMCs under different treatments with or without 10μM EPZ5676. Scale bar = 50μm. Data are means ± sem of 4 samples, *P < 0.05, **P < 0.01, *** P < 0.001, P ≥ 0.05 is not significant (NS)