Validation of microarray results.

<p>Several up-regulated genes at the translational level (translatome) in hypoxic HCT116 cells were validated. RNA isolated from sucrose gradient fractionation was analyzed by quantitative real-time RT-PCR. The distribution of mRNAs in each fraction was calculated and shown as a percentage (%). <b>A</b>. Polysomal profile of β-actin served as a negative control. <b>B</b>. Polysomal profiles of up-regulated genes at both the translational and transcriptional levels (<i>GLUT1</i>, <i>ADM</i>, and <i>VEGFA</i>). <b>C</b>. Polysomal profiles of up-regulated genes at the translational but not transcriptional level (<i>HSPA5</i>, <i>VCAN</i>, and <i>GPR126</i>). <b>D</b>. Translational efficiency of β-actin, GLUT1, ADM, VEGFA, HSPA5, VCAN, and GPR126 mRNAs was calculated and shown as a percentage (%) in HCT116 cells under normoxia and hypoxia. Bar graphs show mean ± standard error from at least three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001).