Fig. 3: 20-HETE is responsible for the ubiquitination and degradation of ACSL4.
From: CYP1B1 inhibits ferroptosis and induces anti-PD-1 resistance by degrading ACSL4 in colorectal cancer
A RKO cells expressing EV or Flag-CYP1B1 were treated with 10 μM AA for 24 h, then analyzed by western blot. B RKO cells were treated with 10 μM 12-HETE or 20-HETE for 24 h and analyzed using western blot. C RKO cells expressing ctrl or CYP1B1 shRNAs were treated with 10 μM 20-HETE for 24 h, followed by western blot analysis. D Viability curves of RKO cells treated with 20-HETE and indicated concentrations of RSL3. E RKO cells were treated with 10 μM 20-HETE and/or sotrastaurin for 24 h, followed by western blot analysis. F, G FBXO10 expression was analyzed using western blot in RKO cells with CYP1B1 overexpression or knockdown. H RKO cells were transfected with Flag-ACSL4, GFP-CYP1B1 and HA-Ub and treated with 10 μM 20-HETE and/or sotrastaurin for 24 h, and 5 μM MG132 for 6 h before harvesting. Thereafter, poly-ubiquitinated ACSL4 was analyzed using western blot.
