Fig. 3.

HIF-1-dependent miR-155 expression in hypoxia. (A) Bioinformatic analysis using the Genomatix software tool MatInspector reveals the presence of three HRE consensus motifs and two NRE consensus motifs in the promoter of the miR-155 gene. (B) Effective siRNA-dependent knockdown of HIF-1α, HIF-2α, and p65 was confirmed by immunoblot analysis. (C) Caco-2 cells were transfected with HIF-1α, HIF-2α, or p65-siRNA prior to hypoxic exposure (48 h), and mature miR-155 expression was analyzed by real-time PCR. (D) Protein levels of HIFα subunits in HIF-1α or HIF-2α knockdown HepG2 cells were analyzed by immunoblotting. (E) HepG2 cells stably expressing shRNA directed against HIF-1α or HIF-2α were exposed to hypoxia, and pri-miR-155 expression was measured by using real-time PCR. n = four independent experiments throughout. Values are presented as means ± the SEM (*, P < 0.05; **, P < 0.01).