Figure 1
From: Hypoxia-inducible miR-182 enhances HIF1α signaling via targeting PHD2 and FIH1 in prostate cancer
miR-183-96-182 cluster is hypoxia-responsive and regulated by HIF1α.
(a,b) The expression of miR-183-96-182 was assayed by RT-PCR in PC-3 cells after exposure to 1% O2 hypoxic environment for 24 hours (a) or treated with hypoxia inducer DFO for 12 or 48 hours (b). (c) DU145 and PC-3 cells were transfected with HIF1α-DM or control empty plasmid. After 48 hours, the cells were harvested and HIF1α protein was determined. Tubulin was used as a loading control. (d,e) Relative expression of miR-183-96-182 was determined in DU145 (d) or PC-3 (e) cells after transfected with HIF1α-DM as indicated. VEGF mRNA was determined as a positive control. (f) C57BL/6 mice were treated with DMOG or vehicle as described in Materials and Methods. The prostate tissues were harvested and the miRNA expression was determined by RT-PCR analysis (n = 3). (g) PC-3 cells were transfected with control or HIF1α siRNA oligos. After 24 hours the cells were exposed to 1% O2 for 24 hours, then the cells were harvested and HIF1α protein was determined. (h) PC-3 cells were transfected with HIF1α siRNA oligos and exposed to hypoxia. The expression of miR-183-96-182 was determined by RT-PCR. miR-210 expression was also determined. Data are mean ± SEM of three independent experiments. *p < 0.05, **p < 0.01. Full-length blots are presented in Supplementary Fig. S6.
