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. Author manuscript; available in PMC: 2017 Feb 25.
Published in final edited form as: Science. 2016 Aug 26;353(6302):929–932. doi: 10.1126/science.aad5755

Fig. 4. Disruption of Akt proline-hydroxylation events leads to a sustained Akt kinase activation and increased colony formation and tumor growth.

Fig. 4

(A to C) IB analysis of WCL derived fromDLD1-AKT1/2−/− cells (A) infected with lentiviruses encoding WT- or P125/313A-Akt1. (B) Cells were deprived of serum for 24 hours followed by stimulation with insulin (0.1 µM), and relative pT308-Akt intensity was quantified in (C). (D to G) Colony formation (D) and soft agar (F) assays were performed with DLD1-AKT1/2−/− cells generated in (A), and were quantified in (E) and (G) (mean ± SD, n = 3 wells per group). *P < 0.05 (Student’s t test). (H and I) Mouse xenograft experiments were performed with the cells generated in (A). Tumor growth curve (H) and tumor weight (I) were calculated (mean ± SD, n = 6 mice per group). *P < 0.05 (one-way analysis of variance test). (J) A schematic representation of cancer patient-associated Akt mutations. (K and L) Glutathione S-transferase pull-down and IP analysis of WCL derived from HEK293 cells transfected with indicated constructs. (M and N) Relative colony numbers were quantified for colony formation (M) and soft agar (N) assays (mean ± SD, n = 3 wells per group). *P < 0.05, **P < 0.01 (Student’s t test).