Figure 3
From: Notch1 haploinsufficiency in mice accelerates adipogenesis
Notch1 haploinsufficiency increases adipogenic transcriptional factors in white adipose tissue. Notch1 expression in adipose tissue was decreased by as much as 50% in N1+/− mice. Notch1 expression in adipose tissue reached a peak value in 8-week-old mice in parallel with fat accumulation especially under HF/HS-fed condition, but such increase was blunted in N1+/− mice. The expression levels of Hes-1 and Pref-1, and Sox9 were decreased in adipose tissues of N1+/− mice. Downregulation of Sox9 enhanced the expression of the master activators of adipogenesis, PPARγ and C/EBPα. (A) Quantitative analysis of Notch1, Notch2, Notch3, and Notch4 mRNA expression levels in adipose tissues of 8-week-old WT and N1+/− mice. (B) Quantitative analysis of Notch1 mRNA expression levels in adipose tissues of 4-, 8-, and 16-week-old mice. (C–G) Quantitative analysis of mRNA expression levels of Hes-1 (C), Pref-1 (D), Sox9 (E), PPARγ (F), and C/EBPα (G) in adipose tissues of 8-week-old mice. (H) Immunoblotting showing cleaved-Notch1 in adipose tissues of 4-week-old mice. Representative western blot of fractionated samples. The cropped blots are used in the figure. The membranes were cut prior to exposure so that only the portion of gel containing the desired bands would be visualized. The samples of fractionation derive from the same experiment and the blots were processed in parallel. Full-length blots are shown in Supplementary information (Page 6). Quantification of western blot bands. Protein expression was normalized to β-actin. (I) Immunoblotting showing cleaved-Notch1 and Pref-1 in adipose tissues of 8-week-old mice. Representative western blot of fractionated samples. The cropped blots are used in the figure. The membranes were cut prior to exposure so that only the portion of gel containing the desired bands would be visualized. The samples of fractionation derive from the same experiment and the blots were processed in parallel. Full-length blots are shown in Supplementary information (Page 6). Quantification of western blot bands. Protein expression was normalized to β-actin. Data are mean ± SD. **P < 0.01, compared with WT mice. n = 8–10 per group (A–G), n = 5 per group (H and I).
