Fig 1: TSC2-deficient cells show enhanced expression of genes linked to ferroptosis. Transcriptomic data comparing (a) Tsc2−/− versus Tsc2+/+ MEF (n = 3) and (b) TSC2(−) versus TSC2(+) AML cells (n = 6), where ferroptosis-linked genes are shown in volcano plots. Thresholds were set at < and >2-fold changes (for down- and upregulated genes, respectively) with an adjusted p-value < 0.05. (c–e) Western blot analysis of ferroptosis associated proteins were also carried out on TfR1, GPX4, HMOX1 and FSP1 (depending on antibody specificity) in TSC cell line models (Tsc2+/+ and Tsc2−/− MEFs, ELT3-T3 (Tsc2+) and ELT3-V3 (Tsc2−), and TSC2(−) versus TSC2(+) AML cells, respectively) treated with either DMSO or 200 nM RSL3 for 4 h, as indicated (3 biological repeats of each shown). TSC2 and β-actin were analyzed as controls. The uncropped bolts are shown in Supplementary Materials.