Figure 6.

The effect of DNA methylation on the gene expression during development and under various diets. C57BL/6J males were fed with a STD from weaning to 8 weeks‐of‐age, and then one group was fed a high fat diet (HF) until 12 weeks‐of‐age (A–D). Body composition of mice measured at 10 days, 8 and 12 weeks‐of‐age. Data are the mean ± SD (error bars) with n = 15, 4, and 4 for 10 days, 8 weeks, and 12 weeks‐of‐age, respectively (E). Relative mRNA levels (in 60 ng total RNA) of Sfrp5 were determined in whole ING fat tissues examined by real‐time PCR. Data are presented as the mean ± SEM (error bars) with n = 3, 4, and 4 at 10 days, 8 weeks, and 12 weeks‐of‐age, respectively. Three duplicates of individual groups are shown. In the 10‐day‐old group, each n contained a pool of ING tissue from five animals (F). Mean percentage values of methylation of the 46 CpG's islands in the promoter region of Sfpr5 gene in the MAF and the stromal vascular fraction (SVF) from ING fat tissue. Data are the mean ± SEM (error bars) with n = 30, 40, and 40 clones for 10 days, 8 weeks, and 12 weeks‐of‐age, respectively (G). Comparison of methylation pattern of DNA isolated from SVF and MAF of ING fat collected from 10 day (n = 3), 8 week (n = 4), and 12‐week (n = 4)‐old mice (H). Sodium bisulfite sequencing analysis of DNA from cellular fractions (MAF, SVF) isolated from ING fat collected from animals at different ages, 10 days (n = 3), 8 weeks (n = 4), and 12 weeks (n = 4). Data are the means only. Differences between groups were analyzed for statistical significance by one‐way ANOVA with multiple comparisons, significance levels indicated by *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001. MAF, mature adipocyte fraction; STD, standard chow diet; SVF, stromal vascular fraction.