Figure 5.

Ad-KO mice fed HFD have alterations in adipocyte ACSL activity, n-6 PUFA fatty acyl-CoA formation, and FFA and phospholipid composition. (A) ACSL isoform gene expression in isolated gonadal adipocytes of 20-week old male ACSL4^floxed or Ad-KO mice fed LFD or HFD for 12 weeks RNA expression quantified by real-time PCR relative to Cyclophilin A. n = 5–8 mice per group. *p ≤ 0.05 ACSL4^floxed vs. Ad-KO within diet; **p ≤ 0.0125 ACSL4^floxed vs. Ad-KO within diet; #p ≤ 0.05 LFD vs. HFD within genotype; ##p ≤ 0.0125 LFD vs. HFD within genotype; Student's t-test. (B) Total ACSL enzyme activity in isolated gonadal adipocytes of 20 week old males fed LFD or HFD for 12 weeks. n = 4–6 mice/group *p ≤ 0.05 ACSL4^floxed vs. Ad-KO within diet. Student's t-test. (C) Long-chain acyl-CoA species determined in gonadal adipose tissue of male ACSL4^floxed or Ad-KO mice on HFD for 12 weeks as determined by two-way ANOVA. *p ≤ 0.05 ACSL4^floxed vs. Ad-KO within diet; **p ≤ 0.0125 ACSL4^floxed vs. Ad-KO within diet. n = 7–8 mice/group. (D) Isolated adipocyte FFA species (no species less than 20 carbons have significant differences) as determined by two-way ANOVA. *p ≤ 0.05 ACSL4^floxed vs. Ad-KO within diet; **p ≤ 0.0125 ACSL4^floxed vs. Ad-KO within diet. n = 6–9 mice/group. (E) Fold-change of isolated adipocyte arachidonic acid, DHA, and linoleic acid phospholipid species. Significant differences between groups represented in yellow (p ≤ 0.05) and italicized (p ≤ 0.01) as determined by two-way ANOVA (Black boxes = Species not detected). n = 6–9 mice/group. Data represent mean ± SEM.