HIF-2α- and HIF-1α-dependent translation efficiency (TE) and mRNA expression analysis in hypoxia.

mRNA TE and steady-state levels were measured in hypoxic U87MG cells treated with non-silencing control or HIF-2α-specific or HIF-1α-specific siRNA pools followed by ribosome density fractionation. TE was defined as the ratio of mRNA abundance in the pooled polysomal fraction to that in the free/monosomal fraction for each condition. Steady-state mRNA expression was defined as the aggregate mRNA abundance across all sequenced fractions for a given condition.