Fig. 5.

Podocyte GSK3 loss causes mitotic catastrophe. a Western blot showing robust knock down of GSK3α and GSK3β in lentiviral Cre transduced floxed podocytes (n = 4). Controls of Cre transduced wild-type podocytes (n = 3) and non-cre transduced GSK3 floxed podocytes (n = 3) also shown. See also Supplementary Fig. 5d. b Only 50% of ciGSK3DKO cells survive 7 days after Cre lenti transduction. ANOVA, Tukey’s post hoc test ***p < 0.001, n = 3 experiments. c Volcano plot of proteomic data revealed that at day-3 following lentiviral transduction, GSK3α and GSK3β levels were significantly reduced (unpaired two-tailed t test p < 0.01). Numerous proteins associated with cell cycle entry were up-regulated. d Phase contrast microscopy shows ciGSK3DKO cells have significantly more multinucleate cells than controls at day 5, unpaired two-tailed t test *p < 0.05. Three fields of view per group, n = 3 experiments. e Representative western blots of cipodGSK3DKO at day 5 reveals increased expression of Cyclin B1 and phosphorylation of Cdk1 and Histone 3 when compared with control cells, n = 3 experiments. See also Supplementary Fig. 6f. f Representative western blots of cipodGSK3DKO and control cells at day 7 reveals apoptosis in knockout cells. Increased levels of cleaved Caspase 3 and cleaved PARP shown, n = 4–5 experiments. See also Supplementary Fig. 6g. g Representative immunohistochemistry showing increased PCNA staining in glomerulus of mpodGSK3DKO mice. Scale bar = 25 μm. h Histology showing a mitotic figure an mpodGSK3DKO mouse podocyte. Scale bar top panel = 25 μm; bottom panel = 10 μm. Data are presented as the mean ± SEM