Figure 8.
Figure 8. TX requires the 3′ UTR of HIF-1α to regulate its translation. (A) HeLa–GFP-Ago2 cells transfected (3:1 or 3:2 Fugene/DNA ratio) with a GFP-tagged HIF-1α construct lacking both the 5′ and 3′ UTR (HIF-1α–GFP) and treated with 25 nM TX overnight. (B) HeLa–GFP-Ago2 cells transfected with a HIF-1α 3′ UTR luciferase expression vector either alone or cotransfected with four HIF-targeting miRNAs (miR), four HIF-specific anti-miRs (anti), or a scrambled miRNA-like sequence (Scram). Luciferase activity was measured in untreated or TX-treated cells as indicated. *, P < 0.05. Error bars indicate mean ± SEM. (C) HIF-1α immunoblot of HeLa–GFP-Ago2 cells transfected with HIF-specific anti-miRs (622, 338, 411, and 519b), a combination of all four, or Cy3 control for 24 h followed by overnight 25 nM TX treatment. Transfection of two non-HIF–related anti-miRs (let7a and 545) was used as a control.

TX requires the 3′ UTR of HIF-1α to regulate its translation. (A) HeLa–GFP-Ago2 cells transfected (3:1 or 3:2 Fugene/DNA ratio) with a GFP-tagged HIF-1α construct lacking both the 5′ and 3′ UTR (HIF-1α–GFP) and treated with 25 nM TX overnight. (B) HeLa–GFP-Ago2 cells transfected with a HIF-1α 3′ UTR luciferase expression vector either alone or cotransfected with four HIF-targeting miRNAs (miR), four HIF-specific anti-miRs (anti), or a scrambled miRNA-like sequence (Scram). Luciferase activity was measured in untreated or TX-treated cells as indicated. *, P < 0.05. Error bars indicate mean ± SEM. (C) HIF-1α immunoblot of HeLa–GFP-Ago2 cells transfected with HIF-specific anti-miRs (622, 338, 411, and 519b), a combination of all four, or Cy3 control for 24 h followed by overnight 25 nM TX treatment. Transfection of two non-HIF–related anti-miRs (let7a and 545) was used as a control.

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