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. 2021 Aug 20;43(5):1285–1298. doi: 10.1038/s41401-021-00736-y

Fig. 5. BBR exacerbates SIRT3 activity and inhibits MAPK/NF-κB signalling in the eWAT of HFD-challenged mice.

Fig. 5

a The levels of iNOS, COX-2 and SIRT3 in eWAT were measured (n = 8). b Macrophages were treated with BBR (0‒160 μM) for 24 h. SIRT3, AcK122 MnSOD, MnSOD and Nampt levels were measured (n = 5). *P < 0.05 BBR vs. Control. c The effects of BBR on SIRT3 deacetylation activity were determined (n = 6). #P < 0.05 LPS vs. Control; *P < 0.05 BBR vs. LPS; ▲P < 0.05 AGK7 + BBR + LPS vs. BBR + LPS. d Mitochondrial lysates from macrophages treated with BBR were collected and evaluated for lysine acetylation using an anti-acetyl lysine antibody (AcK). MnSOD were used as a loading control. e Mitochondrial lysate from macrophages treated with cycloheximide (10 μM) for 1 h followed by treatment with BBR (80 μM) for an additional 2 h was analysed by Western blotting. f Cellular CETSA was performed on macrophages at different temperatures. The data were normalized to the mean value of the respective group at 50 °C (n = 5). g Predicted binding conformation of BBR to SIRT3 and interactions between BBR and SIRT3. h The calculated interaction energy between the substrate and protein in BBR-bound and BBR-free SIRT3. The expression levels of MAPKs (i) and NF-κB (j) were determined in eWAT (n = 8). The data represent the means ± SD. #P < 0.05 HFD vs. RD; *P < 0.05 BBR vs. HFD.