Figure 1. MTDs inhibit HIF-1α... | Rockefeller University Press

Figure 1.

$Figure 1. MTDs inhibit HIF-1α translation. (A) Polysome association profile of MCF7 cells after overnight treatment with 25 nM TX, 25 nM vinblastine (VBL), or 25 µM 2ME2 visualized after sucrose gradient centrifugation. The 40, 60, and 80S ribosomal subunits and polysomes were fractionated and monitored with continuous A254 measurements. Representative profiles and microtubule images after each drug treatment are shown. Bar, 10 µm. (B) RNA was extracted from each fraction, and HIF-1α, GAPDH, and p53 expression was quantified by qRT-PCR. The distribution of each mRNA between nontranslating (1_6) and translating (7_12) fractions is plotted (mean ± SEM; n = 3–10; ***, P < 0.001). (C) Immunoblot of HIF-1α and actin from MCF7 cells treated as in A and exposed to normoxia (N) or 4-h hypoxia to visualize HIF-1α protein. Black lines indicate that intervening lanes have been spliced out.$

MTDs inhibit HIF-1α translation. (A) Polysome association profile of MCF7 cells after overnight treatment with 25 nM TX, 25 nM vinblastine (VBL), or 25 µM 2ME2 visualized after sucrose gradient centrifugation. The 40, 60, and 80S ribosomal subunits and polysomes were fractionated and monitored with continuous A₂₅₄ measurements. Representative profiles and microtubule images after each drug treatment are shown. Bar, 10 µm. (B) RNA was extracted from each fraction, and HIF-1α, GAPDH, and p53 expression was quantified by qRT-PCR. The distribution of each mRNA between nontranslating (1_6) and translating (7_12) fractions is plotted (mean ± SEM; n = 3–10; ***, P < 0.001). (C) Immunoblot of HIF-1α and actin from MCF7 cells treated as in A and exposed to normoxia (N) or 4-h hypoxia to visualize HIF-1α protein. Black lines indicate that intervening lanes have been spliced out.