Fig. 2: Phosphorylation of PDK1 inhibits AKT kinase activity and oncogenic functions.

a IB analysis of DLD1-PDK1 knockout cells stably infected with the indicated constructs. Indicated protein were quantified. (mean ± SD, n = 3, P = 0.004, 0.003). b IB analysis of WCL derived from DLD1-PDK1 knockout cells transfected with the indicated Flag-PDK1 constructs that were serum-starved for 12 h and then treated with the insulin (100 nM) for the indicated time periods before collection for IB analysis. c pT308-AKT/AKT1 ratio in b were calculated. d, e Cells generated in a were subjected to colony formation and soft agar assays. (mean ± SD, n = 3, P = 0.003, 0.003, 0.005, 0.002). f, g, h Cells generated in a were subjected to mouse xenograft assays. Tumor sizes were monitored (f P = 0.0002), and dissected tumors were weighed and calculated (g, h, P = 0.0042, 0.0059). The Error bars in the f, h are mean plus or minus SD, n = 5 mice. *P < 0.05, **P < 0.01. i IB analysis of WCL derived from dissected tumor tissues. j pT308-AKT/AKT1 ratio in (i) were calculated. (mean ± SD, n = 3, P = 0.0033, 0.0018), k, l Graphic representation of pS6 staining derived from tumor tissues k, which were further normalized and quantified l. (mean ± SD, n = 5, P = 6.41E-06, 2.29E-05) (t test), scale bar, 50 μm. Similar results were obtained in n ≥ 3 independent experiments in b. Statistical significance was determined by two-tailed Student’s t-test in a, e, h. j, l and by two-way ANOVA in f. *P < 0.05,**P < 0.01. Source data are provided in Source Data files. EV, empty vector. WT, wild type.