Fig. 5: Genetic or pharmacological depletion of EZH2 inhibits H2O2 induced partial EMT of cultured human tubular epithelial cells.

Starved HK2 cells were exposed to H2O2 (0.5 mM) in the presence of 3-DZNeP (5 μM) for 24 h before cell harvesting. A Cell lysates treated with 3-DZNeP were subjected to immunoblotting analysis with antibodies against EZH2, H3K27me3, Histone H3, α-SMA, E-cadherin, and GAPDH. B–E Expression levels of EZH2, H3K27me3, α-SMA, E-cadherin were quantified by densitometry and normalized with GAPDH and Histone H3. F Cell lysates treated with 3-DZNeP were subjected to immunoblotting analysis with antibodies against OAT1, AQP1, ATPase, and GAPDH. G–I Expression levels of OAT1, AQP1, ATPase were quantified by densitometry and normalized with GAPDH. J Photomicrographs showed the immunofluorescent co-staining of EZH2 and α-SMA in the HK2 cells of each group. HK2 cells were transfected with EZH2 siRNA and scrambled siRNA for 6 h, and then incubated with or without H2O2 (0.5 mM) for an additional 24 h before being harvested for analysis. K Cell lysates transfected with EZH2 siRNA were subjected to immunoblotting analysis with antibodies against EZH2, H3K27me3, Histone H3, α-SMA, E-cadherin, and GAPDH. L–O Expression levels of EZH2, H3K27me3, α-SMA, E-cadherin were quantified by densitometry and normalized with GAPDH and Histone H3. P Cell lysates transfected with EZH2 siRNA were subjected to immunoblotting analysis with antibodies against OAT1, AQP1, ATPase, and GAPDH. Q–S Expression levels of OAT1, AQP1, ATPase were quantified by densitometry and normalized with GAPDH. Data were expressed as means ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. N.S., statistically not significant, with the comparisons labeled. All scale bars = 50 μm.