VLCFA analysis in ACBD5 and ACBD4 KO HEK293 cells.A, analysis of levels of C26:0 and C26:0-lysoPC in control, ACBD4 and ACBD5 KO HEK293 cells. B, analysis of C26:0 and C16:0 β-oxidation (as a percentage relative to controls) in ACBD4, ACBD5 KO, and control HEK293 cells. C, D3-C22 loading assay with analysis of D3C26:0, D3C16:0, and the ratio of D3C16:0/D3C26:0 in control ACBD4 and ACBD5 KO HEK293 cells. Data analyzed by two-tailed unpaired t test; ns, not significant; ∗p ≤ 0.0332; ∗∗p ≤ 0.0021; ∗∗∗p ≤ 0.0002; ∗∗∗∗p ≤ 0.0001. Error bars represent SD, with three experiments per condition. KO, CRISPR knock-out.