Figure 2Figure 2
Gch1 overexpression and its downstream metabolites BH4/BH2 rescue from ferroptosis. (A) Survival of MF-dCas9-Gch1 (Gch1 OE) compared to empty vector MF control (control) cells and GCH1-IRES-Puro (GCH1 OE) cells compared to parental HT-1080 cells (parental), respectively, against inducers of cell death: Ferroptosis induced with 2 μM IKE and Gpx4–/– by using 1 μM tamoxifen with 10 μM α-tocopherol (αToc) rescue. Extrinsic apoptosis induced by 20 ng/mL tumor necrosis factor α (TNFα) with 10 μM z-VAD-FMK (zVAD) rescue. Intrinsic apoptosis induced by 20 μM doxorubicin (Doxo), 25 μM etoposide (Etop), and 12.5 μM colchicine (Colch). Necroptosis induced by 1 μg/mL lipopolysaccharide (LPS) cotreatment with 10 μM zVAD with 10 μM necrostatin-1 (Nec-1) rescue. (B) Dose response curves of BH2 and BH4 starting at 200 μM rescuing from ferroptosis in MF control and parental HT-1080 cells induced by 1 μM RSL3, 2 μM IKE, and Gpx4–/– by 1 μM tamoxifen. (C) Effect of BH2 and BH4 on lipid peroxidation induc