Figure 2.

5-aza-dC demethylates the Wnt10a promoter. A, 5-aza-dC induces Wnt10a expression at the early stage of differentiation. B, Methylation at the CpG sites of the Wnt10a promoter is reduced by 5-aza-dC. A, 3T3-L1 preadipocytes were induced to differentiate with a differentiation medium as described in Materials and Methods and were concurrently treated with either PBS or 5-aza-dC at day 1, 1–2, 3–5, and 6–8 as indicated in the figure, and cells were harvested at day 8 of the differentiation for the measurement of gene expression. Gene expression was measured by real-time RT-PCR and normalized to cyclophilin. All data are expressed as mean ± SEM, n = 6; groups labeled with different letters are statistically different from each other. B, 3T3-L1 preadipocytes were induced to differentiate and were treated with either PBS or 5-aza-dC (0.5μM) at day 0–2 of differentiation for 2 days. CpG methylation was measured by pyrosequencing as described in Materials and Methods. All data are expressed as mean ± SEM, n = 4; *, P < .05 vs control. Con, control; AZA, 5-aza-dC.