Fig. 3: ACLY is the critical mTORC2 effector during brown adipocyte differentiation.
From: mTORC2-AKT signaling to ATP-citrate lyase drives brown adipogenesis and de novo lipogenesis
a Western blot showing ablation of ACLY protein and AKT phosphorylation upon brief 4-OHT treatment of Acly-iKOPBA cells. b, c Acly-iKOPBA and Rictor-iKOPBA cells were differentiated and pparγ2 mRNA expression (n = 6 per group) (b) and Oil Red O (ORO) staining of lipids (c) were quantified at day 10 to determine differentiation efficiency. Scale bar represents 50 μm. d, e Representative differentiation rescue experiments showing ORO staining (d) and Pparγ2 mRNA expression (n = 6 per group) (e) in Rictor-iKOPBA cells stably expressing empty (EV), Myc-ACLY-WT, Myc-ACLY-S455A, or Myc-ACLY-455D constructs. Percentage of ORO in d is relative to empty vector control. Scale bar represents 50 μm. f Direct acetyl-CoA measurements of Rictor-iKOPBA cells (n = 9 per group) stably expressing empty (EV), Myc-ACLY-WT, Myc-ACLY-S455A, or Myc-ACLY-455D constructs. Bar graphs represent mean ± SEM. ***p < 0.001. a represents ***p < 0.001 vs control (EV), and b represents ***p < 0.001 vs Rictor-iKOPBA (EV). Statistical significance was calculated by using two-way ANOVA with Sidak’s test multiple comparisons (b, e, f).
