Figure 4. Knockdown of RBM38 enhances HIF1α expression through mRNA translation.

³⁵S-metabolic labeling assay was performed with H1299 (A) or p53^−/− HCT116 (B) cells. Cells were transduced with a lentivirus expressing a control luciferase (Luc) shRNA or RBM38 shRNA, selected by puromycin for 3 d, and then treated with 500 μM CoCl₂ for 3 h, followed by labeling with ³⁵S-methionine and ³⁵S-leucine. Cell lysates were isolated and used for immunoprecipitation with anti-HIF1α (H1α67, Sigma) or non-immune mouse IgG. The samples from immunoprecipitation were separated in 8% SDS/PAGE and the protein signals were captured by autoradiography.