Fig. 6

DNMT1 regulated PTEN methylation in kidney injuries in HBx-HK-2 cells. A DNMT1 and PTEN levels and localization within HBx-HK-2 cells measured through immunofluorescence. Scale bar: 10 μm. B Western blotting for PTEN, E-cadherin, ZO1, Vimentin, and Fibronectin in HBx-HK-2 cells treated with or without inhibition of DNMT1 or PTEN. C and D Concentrations of IL-6 and TNF-α in HBx-HK-2 cells supernatant exposed to siDNMT1 or siPTEN transfection were detected by ELISA. E Western blotting for P65, p-P65, IκBα and p-IκBα in HBx-HK-2 cells treated with or without inhibition of DNMT1 or PTEN. F Representative BSP analysis for evaluating the DNA methylation status of the PTEN promoter region (21 CpG dinucleotides) in HK-2 cells and HBx-HK-2 cells transfected with siNC or siDNMT1. Methylated CpGs are indicated as black dots; unmethylated CpGs are indicated as empty dots. G Western blotting for E-cadherin, ZO1, Vimentin, and Fibronectin in HBx-HK-2 cells exposed to siPTEN or LY294002. H Western blotting for P65, p-P65, IκBα and p-IκBα in HBx-HK-2 cells exposed to siPTEN inhibition or LY294002. Results are expressed as mean ± SD of 3 separate assays. *P < 0.05; **P < 0.01