Extended Data Fig. 5: Ether lipid synthesis is increased relative to diacyl-PLs upon CTP depletion.
From: Peroxisomal-derived ether phospholipids link nucleotides to respirasome assembly
a, Heatmap of mitochondrial PC (p < 0.10, two-sided Student’s t-test). Data from experiment 2 (n = 5 biologically independent samples). Ether lipids are bolded. b, Lipidomic analysis of PC and PE subclass abundances in cells treated with Vidofludimus or Brequinar (min-max box plot, line at mean, n = 3 biologically independent samples, * left to right Vido q = 0.0041, 0.0041, 0.0016, 0.0476, Vido + uri [compared to Vido] q = 0.0025, 0.0023, 0.0019, 0.0002, 0.0025, 0.0005, 0.0025, Breq q = 0.0005, 0.0005, 0.0043, 0.0040, Breq + uri [compared to Breq] q = 0.0011, 0.0011, 0.0436, 0.0277). c, Quantification of plasmenyl/diacyl PC or PE abundances in cells treated with Vidofludimus or Brequinar (n = 3 biologically independent samples, * left to right Vido q = 0.0002, 0.0306, Vido + uri [compared to Vido] q = 0.0005 0.0011, Breq q = 0.0018, 0.0286, Breq + uri [compared to Breq] q = 0.0053, 0.0199). d, 14C-ethanolamine incorporation into PE in cells treated with Brequinar (n = 2 experiments). Lipid fractions were treated with TCA to cleave the ether vinyl bond, resulting in lyso-PE. e, Western blot of CTPS1 from sgCTPS1 cells (n = 1 experiment). f, BN-PAGE of mitochondrial fractions from sgCTPS1 cells in the presence or absence of cytidine (50 µg/ml) (n = 3 experiments). g, BN-PAGE of mitochondrial fractions from Brequinar treated cells in the presence or absence of CTP (200 µM) (n = 1 experiment). In this figure, cells were treated with drug for 48 hours, heatmap row/column groupings are Pearson’s correlations, data are presented as mean values + /- s.e.m., Student’s t-test with a two-stage linear step-up procedure of Benjamini, Krieger and Yekutieli, with Q = 5%. Breq (Brequinar), uri (uridine), and cyt (cytidine).
