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. 2023 Feb 24;9(8):eade7864. doi: 10.1126/sciadv.ade7864

Fig. 3. CL deficiency does not impair capacity for uncoupled respiration.

Fig. 3.

(A) Transmission electron microscopy images of BAT mitochondria from control and CLS-iBKO mice. Scale bars, 2 μM. (B) mtDNA levels normalized to nucDNA in BAT from control and CLS-iBKO mice. n = 9 to 13 per group. (C) Protein abundance of UCP1, ETS subunits, and CS in whole BAT homogenates. (D) CS activity in whole BAT homogenates. n = 4 to 5 per group. (E) Mitochondrial O2 consumption (JO2) in BAT mitochondria from control and CLS-iBKO mice, measured in the presence of 5 mM pyruvate, 0.2 mM malate, 5 mM glutamate, 5 mM succinate, and 2, 20, and 200 μM ADP. n = 5 to 6 per group. (F) ATP production (JATP) in the presence of 5 mM pyruvate, 0.2 mM malate, 5 mM glutamate, 5 mM succinate, and 2, 20, and 200 μM ADP. n = 5 to 6 per group. (G) Mitochondrial coupling efficiency (ATP/O ratio). n = 5 to 6 per group. (H) UCP1-dependent respiration in BAT mitochondria from control and CLS-iBKO mice. Respiration was stimulated by 5 mM pyruvate and 0.2 mM malate, and UCP1 was subsequently inhibited by 4 mM guanosine diphosphate (GDP). n = 8 to 12 per group. (I) Protein abundance of UCP1, ETS subunits, and CS in isolated mitochondria. (J) UCP1-dependent respiration stimulated by 0.2 mM malate, 5 mM carnitine, and 20 μM palmitoyl-CoA. n = 6 per group. (K) UCP1-dependent respiration stimulated by 0.2 mM malate, 5 mM carnitine, and 20 μM palmitoyl-l-carnitine. n = 3 to 5 per group. (L) Abundance of phosphorylated (Ser563 and Ser660) and total hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and diacylglycerol acyltransferase 1 and 2 (DGAT1 and DGAT2) in whole BAT homogenates. (M) Abundance of ryanodine receptor 2 (RyR2), sarco/endoplasmic reticulum Ca2+–adenosine triphosphatase 2 (SERCA2), tissue nonspecific alkaline phosphatase (TNAP), creatine kinase B (CKB), creatine kinase, mitochondrial 2 (CKMT2), and ADP/ATP carrier 1 and 2 (AAC1 and AAC2) in whole BAT homogenates. Data are presented as ±SEM. *P < 0.05.