Fig. 3: Overexpression of Asc-1 in brown (pre-)adipocytes suppresses thermogenic gene expression.

a Asc-1-HA-2a-4F2HC or EGFP overexpressing brown preadipocytes were grown to 100% confluence, and RNA or protein was collected from the day of differentiation start (preadipocyte), and after 8 days of differentiation with (Rosi) or without rosiglitazone (adipocyte) and relative gene expression of Asc-1 measured (n = 3). b Western blot of ASC-1-HA (45 kDa) and β-actin (42 kDa; n = 2). c Relative gene expression of Ppary (n = 3), Ucp1 (n = 3), Pgc1α (n = 3), and Prdm16 (n = 3). d Quantification of PPARy1 and 2 and UCP1 (33 kDa) protein levels relative to β-actin (42 kDa; n = 6, only preadipocyte in Ppary blot of Rosi n = 4). The same preadipocyte samples were used as controls for the different differentiation conditions. Statistics were calculated using two-way ANOVA with Tukey’s or Sidak’s (for a, Pgc1α in c, and Ucp1 and Ppary1–2 for adipocyte in d) or two-way ANOVA mixed-effects analysis (Ppary1–2 for Rosi in d) multiple comparison post hoc test. Data are shown as mean ± SEM.