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. 2024 Nov 5;65(12):100692. doi: 10.1016/j.jlr.2024.100692

Fig. 3.

Fig. 3

Lipid composition and bioenergetics parameters of Huh-7 cells. A: Schematic representation of the fate of exogenous added fatty acids (FFA) in cells. Once entered in the cells, FFA can be converted into esters of SH-CoA and incorporated into complex lipids or β-oxidized. B: Distribution of [1-14C]-PA or [1-14C]-OA in Huh-7 after 4 h of incubation with the labeled substrates. C: Real-time RT-PCR analysis of CER6 in Huh-7 cells treated for 48 h with BSA, 200 μM PA, or 200 μM OA. D: Vitality test for Huh-7 cells treated for 24 h with BSA, 200 μM PA, 3.7 μM Myriocin (Myr), and PA + Myr. E: Basal respiration measurements in Huh-7 cells treated for 24 h with BSA, 200 μM PA, or 200 μM OA. Cells were inserted in the oxygraphic chamber and treated with 2 μg/μl oligomycin, 1 μM CCCP, 15 nM antimycin A, and 1 μM rotenone. Basal = basal cell respiration value; ATP-dependent = basal-oligomycin value (ATP-dep.); Max uncoupling (CCCP) = CCCP value (MaxUN); non-mitochondrial respiration = basal-CCCP value (Non-mit.); Respiratory Reserve = CCCP-antimycin A value (RR). Oligomycin: inhibitor of complex V of mitochondrial oxidative phosphorylation (ATP synthase). CCCP: carbonyl cyanide m-chlorophenylhydrazone, an uncoupler of mitochondrial oxidative phosphorylation; antimycin A: inhibitor of complex III of mitochondrial oxidative phosphorylation. Data are reported as nmol O2/ml/min/106 cells. F: Basal respiration of Huh-7 cells treated for 24 h with BSA, 200 μM PA, or 200 μM OA, in the presence or absence of 5 μM 2-tetradecyl glycidic acid (TDGA). G: β-oxidation in Huh-7 cells treated with BSA, 200 μM PA, or 200 μM OA, in the presence of or absence of 5 μM TDGA. The picture reports the mitochondrial and peroxisomal contribution to the total β-oxidation. H: Hydrogen peroxide (H2O2) production from Huh-7 cells after 24 h with BSA, 200 μM PA, or 200 μM OA. (I, J) Real-time RT-PCR analysis of CPT1 and PPARα in Huh-7 cells 24-hour-treated with BSA, 200 μM PA, or 200 μM OA. Asterisks indicate differences compared to the control, while hashtags among groups. Values are means ± SD. (∗) P < 0.05; (∗∗) P < 0.01; (∗∗∗) P < 0.005; (∗∗∗∗) P < 0.001.