Fig. 7: Inhibition of GSK3β in CDK5RAP2-depleted cells inhibits senescence that is induced upon inhibition of WIP1 activity.
From: CDK5RAP2 loss-of-function causes premature cell senescence via the GSK3β/β-catenin-WIP1 pathway
A BJ cells transfected with CDK5RAP2 siRNA #2 were treated with TWS119 (5 μM in DMSO) and/or GSK2830371 (10 μM in DMSO), a potent inhibitor of WIP1 activity. Cell lysates were then analyzed by SDS-PAGE and immunoblotting for CDK5RAP2, β-catenin, WIP1, phosphoSer15-p53, p53, and actin (left panel). Actin blot was used as a loading control. Representative blots from one of three independent experiments (n = 3) showing similar results are shown. Ratios of levels of β-catenin and WIP1 vs actin and phosphoSer15-p53 vs. p53, and standard deviation for the 3 independent sets of experiments (right panel) were calculated as described for CDK5RAP2 vs. actin in Fig. 1, with values from cells transfected with control siRNA and treated with DMSO normalized to 1.0. *p < 0.05. B Representative images of SA-β-gal staining (upper panel) are from one of three independent experiments (n = 3) showing similar staining patterns. The number of SA-β-gal positive cells (lower panel) was assessed in ~100 cells per treatment group in each of the 3 independent experiments (n = 3). *p < 0.05.
