Figure 5. Acyl-CoA synthetase activity is impaired in hepatic mitochondria of LTKO mice.

(A) The substrates for fatty acid oxidation. (B-E) ADP-stimulated and oligomycin-induced oxygen consumption rate (OCR) of isolated hepatic mitochondria from NCD-fed flox or LTKO mice with various substrates including octanoylcarnitine (B), palmitoylcarnitine (C), palmitoyl-CoA (D), and palmitate with (+) or without (−) coenzyme A (CoA) (E). n =3 mice per genotype and n = 6 replicates per mouse. Each dot indicates the mean value for each mouse. For data in (B), (C), (D) and (E), *p < 0.05, N.S., no significance (two-way ANOVA followed Holm-Sidak’s multiple comparisons test). (F) ¹⁴C-palmitate oxidation rate in the PMP (plasma membrane permeanilizer, 3 nM)-treated primary hepatocytes from flox or LTKO mice with or without CoA. n = 6 replicates per group. (G) Fatty acid oxidation rate in primary hepatocytes from flox or LTKO mice upon 5 μM triascin C treatment (pretreatment for 30 min and during 3 hours of oxidation assay). n = 6 replicates per group. (H) The mRNA levels of ACSL isoforms in primary hepatocytes by ACSL1 siRNA transfection. n = 3 replicates per group. siNC, siRNA for negative control. ***p < 0.001 (Student t test). (I) Fatty acid oxidation rate in siACSL1 transfected primary hepatocytes from flox or LTKO mice. n = 6 replicates per group. For data in (F), (G), and (I), *p < 0.05, **p < 0.01, N.S., no significance, (two-way ANOVA followed Holm-Sidak’s multiple comparisons test). All data in the figure are shown as the mean ± SEM. See also Figure S5.