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. 2017 Dec 18;128(2):625–643. doi: 10.1172/JCI89212

Figure 7. HIF-1α–enhanced Dicer proteolysis in miRNA biogenesis.

Figure 7

(A–C) Effects of HIF-1α on Dicer-dependent miRNA maturation. HCT116 cells overexpressing HIF-1α or HLH domain–truncated HIF-1α (A), knockdown of HIF-1α (B), or restoration of Dicer (C) were subjected to quantitative reverse-transcription PCR (qRT-PCR) for analyzing the expression of miRNAs, including let-7 family members (let-7a, let-7b, let-7d), miR-200b, and miR-451. (A) The ratios of mature to pri-miRNAs for let-7a, let-7b, let-7d, miR-200b, and miR-451 were determined using data sets shown in Supplemental Figure 6, A and B. (D–F) Effects of HIF-1α–regulated Dicer on the expression of miR-200b and let-7b–targeted genes. HCT116 cells were transfected with ZEB1, LIN41, or AURKB 3′ UTR luciferase reporter. The expression of luciferase was assayed in HIF-1α–overexpressing cells with or without Dicer restoration (D). miR-200b (E) or let-7b (F) was restored in HIF-1α–expressing cells. Aurora B, let-7b–targeted gene was analyzed by Western blot. The immunoblots presented were derived from replicate samples run on parallel gels (E). Data are presented as mean ± SD, with at least n = 3 per group. Unpaired, independent groups of 2 were analyzed by 2-tailed Student’s t test. Multigroup comparisons were analyzed by 1-way ANOVA with Tukey’s post hoc test.