Extended Data Fig. 9: AMPK activation promotes the reliance of cancer cells on DHODH-mediated ferroptosis defence.
From: De novo pyrimidine biosynthetic complexes support cancer cell proliferation and ferroptosis defence
a, HeLa cells were treated with A-769662 (200 μM), AICAR (200 μM) or MK-8722 (5 μM) for 12 h. The expression of indicated proteins was detected by immunoblotting. b, HeLa cells were treated with brequinar (0.3 μM) or/and AICAR (40 μM)/A-769662 (200 μM)/MK-8722 (1 μM). Survival of the cells was measured after 3 days of treatment. c, Cell viability of HeLa cells treated with brequinar (0.3 μM) or/and AICAR (40 μM)/A-769662 (200 μM)/MK-8722 (1 μM) for 72 h following pretreatment with vehicle or cell death inhibitors for 24 h. Ferr-1 (10 μM), Z-VAD-FMK (10 μM), necrosulfonamide (2 μM). d,e, Cell viability of H1299 (d) and KYSE-150 (e) cells treated with brequinar (0.3 μM) or/and AICAR (40 μM)/A769662 (200 μM)/MK-8722 (1 μM) for 72 h following pretreatment with vehicle or Ferr-1 (10 μM) for 24 h. f, The body weights of mice with the indicated treatments at the endpoint of experiments. n = 5 (vehicle), n = 5 (AICAR), n = 5 (brequinar), n = 5 (AICAR and brequinar) or n = 6 (vehicle), n = 6 (MK-8722), n = 6 (brequinar), n = 6 (MK-8722 and brequinar). g, HeLa cells were treated with the indicated concentrations of brequinar(BQR) or vidofludimus (Vido). Survival of the cells was measured after 2 days of treatment. h, WT and AMPKα1/2−/− HeLa cells were treated with 2.5 μM brequinar in combination with 0 or 400 μM uridine. Cell survival was measured after 3 days of treatment. (b-h) Values are shown as the mean ± SD, n = 3 (b-e,g,h) biologically independent samples or n = 5 or 6 biologically independent mice (f). P values were obtained using one-way ANOVA with Tukey test (b) and unpaired two-tailed Student’s t-test (c-e, g,h). (a) Data are verified in three replicates with similar results.
