Extended Data Fig. 7: EMCS harbor phospholipids containing PUFAs that are prone to ferroptosis.

a, Representative immunoblot for IP3R3, PERK and CNX from inputs and EMCSs fractions of PERK^WT and PERK^KO MEFs after 6 h RSL3 (0.125 µM). Arrow indicates PERK. b, Orthogonal partial least squares discriminant analyses (OPLS-DA) score plots showing the separation of PERK^WT input (RSL3+Fer-1) and PERK^WT EMCS (RSL3+Fer-1) in two distinct groups. (n = 3 biological replicates). c, Total contents of polyunsaturated fatty acids (PUFAs) in four subclasses of PLs - PUFA-phosphatidylcholine plasmalogen (PCp), PUFA-phosphatidylserine (PS), PUFA-phosphatidylinositol (PI) and PUFA-phosphatidylglycerol (PG) in EMCSs vs input in PERK^WT cells. (n = 3 biological replicates). d, Total contents of cardiolipin (CL) in EMCSs vs input in PERK^WT cells. (n = 3 biological replicates). e, Orthogonal partial least squares discriminant analyses (OPLS-DA) score plots showing the separation of PERK^KO input (RSL3+Fer-1) and PERK^KO EMCS (RSL3+Fer-1) samples in two distinct groups. (n = 3 biological replicates). f, Orthogonal partial least squares discriminant analyses (OPLS-DA) score plots showing the separation of PERK^WT input (RSL3) and PERK^WT input (RSL3+Fer-1) samples in two groups. (n = 3 biological replicates). g, Orthogonal partial least squares discriminant analyses (OPLS-DA) score plot showing the separation of PERK^WT EMCS (RSL3) and PERK^WT EMCSs (RSL3+Fer-1) samples in two groups. (n = 3 biological replicates). h, Volcano plot showing the RSL3-induced changes in the content of oxidized PLs in EMCSs of PERK^WT cells (RSL3 vs RSL3+Fer-1). (n = 3 biological replicates). i, Volcano plot showing the RSL3-induced changes in the content of oxidized PLs in input of PERK^WT cells (RSL3 vs RSL3+Fer-1). (n = 3 biological replicates). In c, d, quantitative data are mean ± SEM. In c, d, statistical significance was determined by two sided unpaired t-test. NS, not significant (P > 0.05), *P ≤ 0.05, **P < 0.01, ***P < 0.001. Source numerical and statistical data are provided.

Source data