HIF-1α mRNA is sequestered to P-bodies upon microtubule disruption. (A) HeLa–GFP-Ago2 cells treated with 25 µM 2ME2 or 25 nM TX overnight, lysed, and immunoprecipitated (IP) using an anti-GFP antibody. Ago2 protein was detected by immunoblotting. Ago2-bound RNA was extracted, and HIF-1α, p53, and GAPDH expression is displayed as mean fold change ± SEM (n = 3; *, P < 0.05). (B) RNA from Ago2 unbound lysates was processed as in A (n = 3; *, P < 0.05). (C) Bar graph showing the percentage of HIF-1α mRNA localized to P-bodies after the indicated drug treatments. (D) HeLa–GFP-Ago2 cells transfected with 100 nM HIF-1α MB (red) and treated with 100 nM TX or 100 µM 2ME2 for 1 h. Bar, 10 µm. Right column shows the percentage of overlap (mean ± SEM; n = 3) between Ago2 foci and HIF-1α MB (***, P < 0.001; Videos 6 and 7).