Figure 5

Notch signaling inhibits expression of Ppargc1a and Prdm16 genes in cultured white adipocyte. (a) Gene expression of Notch1 and Hes1 (left), Prdm16 and Ppargc1a (right) in Notch1^flox/flox preadipocytes after transfection with Cre or GFP plasmids (control). (b) Gene expression of Notch target (left), BAT-related genes (middle) and protein level of N1ICD, UCP1, PGC1-α (right) in cultured WT white adipocytes treated with DAPT during induction and differentiation, n = 6. (c) Gene expression in Rosa^N1ICD preadipocytes after transfection with Cre or GFP (control) plasmids. (d,e) Gene expression of WT preadipocytes after transfection with Hey1 (d) or Hes1 (e) plasmids. (f) Conserved N box domains on Prdm16 (top) and Ppargc1a (bottom) promoters of both human and mouse. (g) Chromatin immunoprecipitation (ChIP) using Hes1 antibody followed by qPCR assay, n = 4. (h) Luciferase assay of HEK293 cells after cotransfection of pGL3-PGC1-α (or pGL3-Basic) plasmids with Renilla plasmids and Hes1 (or its control, ctrl) plasmids, n = 5. *P < 0.05 and **P < 0.01. Data are means ± SEM. n = 3 unless otherwise indicated.