Fig. 3: Mutation of W80 on AKT1 confers resistance to MK-2206 but not ipatasertib.

a Par or M-R7 cells were either not-transfected (N/T) or transfected with siControl or siRNA targeting AKT1 (siAKT1) and grown overnight. Cells were then treated with MK-2206 and viability was assessed 4 days later (left). Levels of indicated proteins were assessed by immunoblot 6 days following transfection (right). b Allele frequencies of the AKT1 W80C mutation in individual cell lines, as detected by exome-seq. c LNCaP Par, M-R7 or Par cells stably overexpressing cumate-inducible, siRNA-resistant AKT1 WT or W80C, or EV were treated with 10 μg/ml cumate and 4 days later, re-plated in 10 μg/ml cumate. The following day, cells were treated with a dose range of MK-2206 and viability was assessed after a further 4 days. d Response of M-R7 cells stably overexpressing cumate-inducible siRNA-resistant AKT1 WT, AKT1 W80C, or EV to MK-2206 was assessed as in c. e As in d except M-R7 EV, AKT1 WT and AKT1 W80C lines were transfected with siAKT1 when re-plating in 10 μg/ml cumate. f As in e except the response of cells to ipatasertib was assessed. g Response of Ba/F3 cells simultaneously overexpressing MEK1 N3 and AKT1 WT, E17K or W80R to MK-2206 (left) or ipatasertib (right) was assessed 4 days after plating cells in the absence of IL-3 with the viability assay. Error bars represent standard error of the mean (SEM); n = 4 replicates in a, c–g. See also Supplementary Fig. 5 and Supplementary Dataset 3, 4. Source data are provided as a Source Data file.