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Fig. 6 | Molecular Cancer

Fig. 6

From: SPOP-mediated ubiquitination and degradation of PDK1 suppresses AKT kinase activity and oncogenic functions

Fig. 6

Patient-derived PDK1 mutants adjacent degron block SPOP-mediated PDK1 degradation. A A schematic presentation of the patients-associated PDK1 mutations. B IB analysis of WCL and GST-pulldown products derived from HEK293T cells transfected with indicated constructs. C IB analysis of WCL derived from HEK293T cells transfected with Flag-PDK1 (WT, Q387R, Q387H) and with/without HA-SPOP. D-E IB analysis WCL derived from of HEK293T cells transfected with indicated constructs. Where indicated, cells were treated with CHX (100 μg/ml) for the indicated time points before harvested. PDK1 protein abundance in (D) was normalized and quantified (E) (mean ± SD, n = 3) (t test), *P < 0.05. F–H IB analysis of DLD1-PDK1 knockout cells stably infected with indicated constructs (F). Resulting cells were subjected to colony formation and soft agar assays (G), and the relative colony numbers were quantified (H). (mean ± SD, n = 3) (t test), **P < 0.01. I-L Cells generated in (F) were subjected to mouse xenograft assays. Tumor sizes were monitored (I), and dissected tumors were weighed (J, K) and subjected for IB analysis (L). (mean ± SD, n = 8) (ANOVA test for I, and t test for K), *P < 0.05., **P < 0.01

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