Fig. 4

Lnc-PDZD7 upregulates EZH2 by competitively binding miR-101 in HCC. a, The gene expression were analyzed by the mRNA microarray in Lnc-PDZD7 down-regulated HepG2 cells. Top 10 downregulated genes were shown in the heatmap. b, The protein level of EZH2 was detected by Western blotting in siLnc-PDZD7 or siLnc-scr transfected HepG2 cells and Lnc-PDZD7 or Lnc-vec transfected Bel-7402 cells. ß-Actin was used as a loading control. c, The expression of miR-101 was determined using qRT-PCR in cells transfected with siLnc-PDZD7 or Lnc-PDZD7. d, Binding sites of miR-101 in the 3’UTR of Lnc-PDZD7 (Lnc-PDZD7 wt) was predicted using bioinformatics prediction tools. The mutated site in the 3’UTR of Lnc-PDZD7 is shown as Lnc-PDZD7 mut. e, The relative luciferase activities were analyzed in HepG2 cells co-transfected with miR-101 mimics or miR-NC and luciferase reporter vectors psiCHECK2- Lnc-PDZD7-WT or psiCHECK2- Lnc-PDZD7-Mut. f, The protein expression were examined by Western blotting in EZH2 plasmid or EZH2-vec transfected Lnc-PDZD7-depleted HepG2 cells. g, The protein expression were examined by Western blotting in shEZH2 or shEZH2scr transfected Lnc-PDZD7-overexpressing Bel-7402 cells. The data are shown as the means ± S.D. of three replicates (*P < 0.05)