Loss of FASN decreases palmitate levels in Pink1 deficiency. (A and B) Free fatty acid levels are reduced in pink1^B9 mutant flies and are further reduced by heterozygosity of dFasn2 in pink1^B9 mutant flies (pink; A) or by Cerulenin (green) in pink1^B9 mutants (B) compared with pink1^RV control flies (black; n = 5 assays). (C and D) Free fatty acid levels (C) and palmitate levels (D) in Pink1^−/− MEFs incubated with Cerulenin (green) are reduced compared with those not incubated with Cerulenin (black) and to Pink1^+/+ control MEFs. n = 5 assays. (E–J) Quantification (E, G, and I) and images (F, H, and J) of protein levels of FASN normalized to actin in Pink1^−/− MEFs (E and F), patient-derived fibroblasts (G and H), and H₂O₂-treated wild-type MEFs (I and J). n = 3 assays. Bonferri’s analysis of variance (A–D) and Student’s t test (E, G, and I): *, P < 0.05; **, P < 0.01; ns, not significant. Data represent percentages (A–D) or normalized means (E, G, and I) ± SEM.