Fig. 4: Elovl1-deficient CD8+ T cells show a rewired lipid profile and an increase in cholesterol levels.
a, Schematics of ELOVL family activity. b,c, LS-MS lipidomics on in vitro sgNT and sgElovl1 OT-I T cells (n = 3). Heatmap representing the relative abundance of each specific lipids (b) and quantification of total cholesterol (c). d, Total cellular cholesterol quantification via Amplex Red cholesterol kit in in vitro sgNT or sgElovl1 OT-I T cells (n = 3). e, Membrane cholesterol quantification with FILIPIN III (n = 4). Representative histogram (left) and MFI quantification (right) in in vitro sgNT or sgElovl1 OT-I T cells. f, Flow cytometry quantification of plasma membrane cholesterol with AlexaFluor 647 conjugated ALOD4 in cultured sgNT and sgElovl1 OT-I T cells (n = 3). Representative histogram (left) and quantification (right). g, Flow cytometry quantification of the membrane fluidity using the fluorescent probe PDA in vitro in sgNT or sgElovl1 OT-I T cells. Data are shown as excimer/monomer ratio (450 nm/400 nm) (n = 4). h, Membrane cholesterol quantification with FILIPIN III (n = 5) in sgNT or sgElovl1 OT-I T cells infiltrating KPC_OVA tumours of mice treated with anti-PD-1. Data are presented as the mean ± s.e.m. Statistical significance was assessed by two-tailed unpaired (c,e,f,h) and paired (g,d) Student’s t-test.
