Figure 5. Sirt1 decreased brown adipocytes apoptosis and reduced ATF4 level.

Brown adipocytes were pre-infected with recombinant vectors of Sirt1 (pAd-Sirt1 or si-Sirt1) for 48 h. n = 3 for each treatment. (A) Protein level of Sirt1 and Sirt2 in brown adipocytes; (B) ATP level of brown adipocytes; (C) Cytosolic Ca²⁺ concentration and Flow cytometry (FCM) analysis of Cytosolic Ca²⁺ in brown adipocytes; (D) Annexin V-FITC/PI double staining and flow cytometry analysis of brown adipocyte apoptosis stages; (E) Protein levels of GRP78, Smad3, Chop, Bcl-2, Bax, Apaf-1, Cleaved-Caspase9 and Cleaved-Caspasw3 in brown adipocytes. pAd-Sirt1: recombinant adenovirus over-expression vector of Sirt1, si-Sirt1: recombinant lentiviral interference vector of Sirt1. Values are means ± SEM. *p < 0.05 compared with the control group.