Journal: Protein & Cell

Article Title: Proteomic analysis of ferroptosis pathways reveals a role of CEPT1 in suppressing ferroptosis

doi: 10.1093/procel/pwae004

Figure Lengend Snippet: Ferroptosis inhibition by CEPT1 is partially dependent on phospholipases. (A–D) Pulldown assays were performed with S protein beads (SFB-tagged proteins were used as the baits), and the indicated proteins were detected by WB. (E) Cell death was measured by propidium iodide (PI) staining in control (sgControl), DDHD1 -knockout, or DDHD2 -knockout (sg1/2) 786-O cells overexpressing CEPT1 or transfected with empty vector (EV) treated with 50 nmol/L RSL3 for 24 h. (F) Cell death was measured by PI staining in control (shControl) and ABHD3 -knockdown (sh1/3) 786-O cells overexpressing CEPT1 or transfected with EV treated with 50 nmol/L RSL3 for 24 h. (G and H) Cell death was measured by PI staining in control (sgControl) and PLAA -knockout (sg1/2) 786-O cells overexpressing CEPT1 or transfected with EV treated with 50 nmol/L RSL3 (G) or 10 µmol/L erastin (H) for 24 h. (I and J) Cell death was measured by PI staining in 786-O cells overexpressing CEPT1 or transfected with EV treated with DMSO, 50 nmol/L RSL3 (I), or 10 µmol/L erastin (J) with 1 µmol/L pyrrophenone (PY) or indicated combinations for 24 h. (K) PLA2 enzyme activities were measured in 786-O cells overexpressing CEPT1 WT or indicated mutants or transfected with an empty vector (EV).

Article Snippet: Ferroptosis inducer (1S,3R)-RSL3 (#19288), erastin (#17754), arachidonic acid (#90010), and pyrrophenone (#13294) were from Cayman Chemical.

Techniques: Inhibition, Staining, Control, Knock-Out, Transfection, Plasmid Preparation, Knockdown