Figure EV4. De‐repression of the β‐catenin‐mediated translation repression by Wnt stimulation is independent of transcription.

A10 cells were serum deprived overnight prior to 1 h pre‐treatment with either actinomycin D (0.5 μg/ml) or its solvent (DMSO), and the cells were stimulated with Wnt‐3a (100 ng/ml) or its solvent (0.1% BSA in PBS) for 4 h. Before harvesting for Western blot analysis, the cells were further treated with 0.5 μM puromycin for 15 min. Total β‐catenin and phospho‐β‐catenin (S33/37/T41) levels indicate effective Wnt‐3a stimulation. P53 was used as a control for actinomycin D activity, and actin was used as a loading control. The average puromycin/actin ratio was graphed (n = 3). Error bars indicate standard deviation. An independent samples t‐test was performed (two‐tailed), with the P‐value indicated above the error bar.Source data are available online for this figure.