Figure S3 from GSK3β-Mediated Expression of CUG-Translated WT1 Is Critical for Tumor Progression

A cycloheximide (CHX) chase assay was used to detect turnover of cugWT1 proteins. HeLa cells were transfected with V5-cugWT1 wt or mutants (S64A, S68E) with Flag-FBXW4 or Flag-FBXW5 and then treated with CHX (20 μg/mL) to prevent further protein synthesis. Whole cell lysates were harvested and prepared at 0, 3, and 6 h post CHX treatment and visualized by Western blotting.