1. 1 ++++ ---- Dox E17K WT Basal mediumGrowth medium p-AKT S473 FLAG PI3K  p-AKT T308 p-mTOR S2448 p-mTOR S2481 mTOR Actin Davies BR et al. Supplementary Figure 1 MCF10-AKT1 E17K and MCF10A-AKT1 WT cells were cultured in basal or growth medium in the presence or absence of dox, and proteins were detected by Western blotting.

2. 2 -Dox+Dox Davies BR et al. Supplementary Figure 2 AKT1 E17K AKT1 WT HER2 V659E RFP AKT1 E17K AKT1 WT HER2 V659E RFP Triplicate wells are shown from colony formation assay in soft agar.

3. 3 Davies BR et al. Supplementary Figure 3 AKT1 E17K -DoxAKT1 E17K +Dox AKT1 WT -DoxAKT1 WT +Dox RFP +DoxRFP -Dox Mean colony diameter (  M) -Dox+Dox AKT1 E17K 101226 AKT1 WT 9597 RFP9196 Mean number of cells per colony -Dox+Dox AKT1 E17K 71479 AKT1 WT 6164 RFP6763 Number of colonies of particular diameters, mean colony diameter and mean number of cells per colony are shown from the soft agar assay.

4. 4 Genomic DNA HBCx2 DNA Davies BR et al. Supplementary Figure 4 Detection of AKT1 E17K mutation in the HBCx2 explant model

5. GI50 = 2.4  MGI50 = 1.6  M Davies BR et al. Supplementary Figure 5 Effect of AKT inhibitors on signaling proteins and growth of MGH-U3 cells