Fig. 5: MUFA-linked ether phospholipids promote the assembly of mitochondrial super-complexes.

a, b BN-PAGE showed mitochondrial supercomplexes (SC) in sensitive (PATC66/108) and resistant (PATC124/148) cells (a). Quantifications of the high molecular weight supercomplex (hwmSC) are shown in (b). Data represent mean ± S.D of three biologically independent replicates. c, d Mitochondrial supercomplexes as shown with BN-PAGE (c) and quantifications of hwmSCs (d) in sensitive (PATC66/108) cells grown in the presence or absence of 100 μM ether-MUFA (O-C16-18:1 phosphatidylcholine (PC)) for 24 hr. Data represent mean ± S.D of 4 biologically independent replicates. e, f Mitochondrial supercomplexes as shown with BN-PAGE (e) and quantifications of hwmSCs (f) in sensitive (PATC66/108) cells grown in the presence or absence of 100 μM ether-PUFA (O-C16-20:3 PC) for 24 hr. Data represent mean ± S.D of three biologically independent replicates. g, h Mitochondrial supercomplexes as shown with BN-PAGE (g) and quantifications of hwmSCs (h) in PATC124 cells infected with control sgRNA (sgCTRL) or sgRNA targeting GNPAT(sgGNPAT) in the presence or absence of 100 μM O-C16-18:1 PC or 100 μM ether-PUFA (O-C16-20:3 PC) for 24 hr. Complex V subunit protein APT5A was detected by SDS-PAGE western blotting as control protein. Data represent mean ± S.D of 5 biologically independent replicates. i Schematic representation of the major conclusions. Peroxisome-derived ether phospholipids, especially those linked with MUFAs, enhance mitochondrial ETC supercomplexes assembly to maintain redox balance and promote resistance to mitochondrial complex I inhibition. Impairment of MUFAs-linked ether phospholipids synthesis by targeting SCD1, GNPAT, or APGS synergizes with mitochondrial complex I inhibition to induce mitochondrial ROS and cell death. Statistical analysis by two-tailed Students’ unpaired t test with significance indicated (b, d, f, h). Source data are provided as a Source Data file.