Extended Data Fig. 7: Investigation of GPX4 degradation mechanism after treatment with ferroptosis inducers.
From: Characterization of a patient-derived variant of GPX4 for precision therapy
a, HT1080 overexpressing exogenous WT or R152H GFP-GPX4 and a control line were tested for FIN56 sensitivity. Data are plotted as means ± SD, n = 3 biologically independent samples. b, Western blot of HT1080 OE GFP-GPX4WT and HT1080 OE GFP-GPX4R152H after treatment with ferroptosis inducers with GPX4 and GAPDH antibodies, with lanes arranged for cell line comparison. Triplicate experiments were repeated independently with similar results, which were shown in Extended Data Fig. 7c,e,f. c, Western blot of HT1080 OE GFP-GPX4WT and HT1080 OE GFP-GPX4R152H after treatment with ferroptosis inducers with GPX4 and GAPDH antibodies, with lanes arranged for ferroptosis inducer comparison. Triplicate experiments were repeated independently with similar results, which were shown in Extended Data Fig. 7b,e,f. d, The endogenous GPX4 in HT1080 OE GFP-R152H-GPX4 were tested for vulnerability to the degradation induced by RSL3, ML162, FIN56, and IKE. Data are plotted as means with range of two biologically independent experiments. The corresponding blots are shown in Extended Data Fig. 7b,c. e, Western blot of HT1080 OE GFP-GPX4WT after treatment with ferroptosis inducers with GPX4 and GAPDH antibodies. Triplicate experiments were repeated independently with similar results, which were shown in Extended Data Fig. 7b,c. f, Western blot of HT1080 OE GFP-GPX4R152H after treatment with ferroptosis inducers using GPX4 and GAPDH antibodies. Triplicate experiments were repeated independently with similar results, which were shown in Extended Data Fig. 7b,c.
