Fig. 2. ACSL4-dependent PL remodeling is required for ferroptosis resistance of TRCs.
a, The radar chart illustrates changes of SFA-PLs, MUFA-PLs and PUFA-PLs in HONE1 TRCs and bulk tumor cells (left). The bar chart (right) represents the relative increment of these PLs. b, The content of esterified AA (C20:4) or AdA (C22:4) PE or PC molecular species in HONE1 TRCs and bulk cells. c, The formation rate of AA-d8-CoA in reactions catalyzed by ACSL4 purified from HONE1 TRCs and bulk cells at different timepoints. AA-d8 and coenzyme A were used as substrates. AA-d8-CoA was undetectable in the samples with no enzyme added. d, The content of esterified AA (C20:4) or AdA (C22:4) PE molecular species in TRCs and bulk tumor cells from ACSL4-knockout HONE1 cells and parental cells. e, The content of esterified AA (C20:4) or AdA (C22:4) PC molecular species in TRCs and bulk tumor cells from ACSL4-knockout HONE1 cells and parental cells. f,g, TRCs and bulk cells from ACSL4-knockout HONE1 cells (sg ACSL4) or parental cells (sg GFP) were treated with RSL3 (15 µM) for 13 h. The content of ferroptotic cell death signal PE (38:4)-OOH (f) and PE (40:4)-OOH (g) was measured by LC–MS/MS. P = 0.0000596 and 0.5509789. h, TRCs and bulk tumor cells from ACSL4-knockout HONE1 cells or parental cells were treated with RSL3 (15 µM) or FIN56 (10 µM). The percentage of dead cells was measured after indicated treatment for 16 h. Data are shown as mean ± s.d.; unpaired two-tailed t-test (b,c,h) or one-way ANOVA (d–g). n = 3 independent experiments. NS, not significant.