Supplementary Figure 4: Nrf2 is necessary for AKT2(E17K)-mediated stimulation of GSH biosynthesis and resistance to oxidative stress.

NRF2 was knocked down over 72 h. (a) Nrf2 knock-down was confirmed by immunoblot analysis (data is representative of three independent experiments). (b) Total glutathione levels in serum-starved cells (data are from one experiment that was independently repeated two times with similar results (Supplementary Table 1)). (c) Serum-starved cells were treated with 500 μM H₂O₂ for 4 h (data is representative of three independent experiments). (d) Distribution of RPPA Z-scores for Akt S473 phosphorylation in patient tumors from TCGA BRCA data set. Breast tumors with Akt pS473 levels greater than 2 standard deviations from the mean were classified as ‘Akt pS473 high’, while tumors with Akt pS473 levels less than 1 standard deviation from the mean were classified as ‘Akt pS473 low’. Unprocessed original scans of blots are shown in Supplementary Fig. 6.