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. 2020 Jul 9;36:101633. doi: 10.1016/j.redox.2020.101633

Fig. 4.

Fig. 4

Limiting amino acids enhances Atgl-mediated fatty acid oxidation in beige adipocytes.

A. Single gene expression analysis in primary beige adipocytes cultured in a medium poor in amino acids (AAR) or adrenergically stimulated by CL316,243 (CL). Data shown are the result of 3 separate experiments. Data are presented as mean ± S.D. ***p < 0.001 AAR or CL vs Ctr. B-D. Venn diagram of up-regulated gene transcripts and proteins in sWAT isolated from adult male mice fed with LPHC diet (n = 3 mice) for 2 weeks (B). Functional enrichment analysis of overlapped genes (n = 61) and the relative GO terms for biological processes are reported (C). Schematic representation of key metabolic enzymes over-represented in sWAT (n = 61) related to fatty acid oxidation, TCA cycle, ETC and glycolysis pathways. For each enzyme, the corresponding LPHC-induced changes in either mRNA or protein concentration were reported as a heatmap (D). E. Seahorse analysis of X9 beige adipocytes cultured in a medium poor in amino acids (AAR) supplemented with Atgl inhibitor Atglistatin (ATGLi) or vehicle. The oxygen consumption rate (OCR) (pmol/min) was monitored and maximal respiration is reported in the bar graphs. Reported data are the result of 3 separate experiments. Data are presented as mean ± S.D. **p < 0.01 ATGLi vs vehicle. F. Mitochondrial proton gradient (Δp) was measured by cytofluorimetry after staining with MitoTracker Red CMXRos in X9 beige adipocytes cultured in a medium poor in amino acids (AAR) supplemented with Atgl inhibitor Atglistatin (ATGLi) or vehicle. Reported data are the result of 3 separate experiments. Data are presented as mean ± S.D. **p < 0.01 ATGLi vs vehicle. G. Schematic representation of FAPs isolated from WT or AtglKO mice. Mitochondrial proton gradient (Δp) and basal oxygen consumption were measured. Reported data are the result of 3 separate experiments. Data are presented as mean ± S.D. *p < 0.05; ***p < 0.001 AtglKO vs WT (Student t-test). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article).