Fig. 3. miR-16 repressed HIF-1α via a targeted intervention of 14-3-3η.
a Potential miRNAs of 14-3-3η as predicted by TargetScan, and the target sequences of miR-16 in the 3′-UTR of 14-3-3η mRNA. b qPCR analysis in triplicate of the expression of miR-16 in HuH7 and HuH7SR cells. c (Left) HuH7SR cells were co-transfected by scrambled or miR-16 mimic in the presence of pGL3-14-3-3η 3′-UTR (WT or MT)-Luc constructs. Luciferase reporter assay analysis in triplicate of the effects of miR-16 on 14-3-3η-3′-UTR. HuH7SR cells were transfected by scrambled or miR-16 mimic (middle), while HuH7 cells were transfected by scrambled or anti-miR-16 (right), after then, the expression of 14-3-3η mRNA was determined in triplicate via qPCR. d–f HuH7SR cells were transfected by scrambled, miR-16 mimic, or miR-16 mimic plus 14-3-3η plasmid. d IB analysis of the expressions of 14-3-3η and HIF-1α with different treatments. e IP analysis of the ubiquitination of HIF-1α. f Immunostaining analysis of the expression and intracellular distribution of HIF-1α