Figure 3.

HNF4G is a downstream target of miR-34a in bladder cancer. (a) A predicted seed region (red) in the 3′-UTR of HNF4G was shown (top). The mutated sequence used was highlighted in blue (bottom). (b) HEK293T cells were cotransfected with either 50 nM miR-34a mimics or NC oligos and 200 ng plasmid carrying either Wt or Mut 3′-UTR of HNF4G. The relative firefly luciferase activity normalized with Renilla luciferase was measured 48 h after transfection. (c) Western blot analysis showed that the HNF4G protein levels in bladder cancer cells were inversely correlated with miR-34a expression as in Figure 1. (d) Knockdown of endogenous miR-34a remarkably increased HNF4G expression at protein level, while ectopic overexpression significantly decreased HNF4G expression in bladder cancer cells. ^* P < 0.05. Error bars depict s.e.m.