Figure 2
From: Inhibiting DNA methylation switches adipogenesis to osteoblastogenesis by activating Wnt10a
The Wnt10a gene is demethylated and up-regulated by 5-Aza-dC in 3T3-L1 cells.
(A) Real-time PCR screening on repressor gene expression of 3T3-L1 cells treated with 5-Aza-dC during adipogenesis. (B) Wnt10a mRNA expression is dramatically induced by early treatment of 5-Aza-dC. (C) Pyrosequencing analysis showed that 5-Aza-dC treatment significantly reduces DNA methylation levels on all the 33 CpG sites measured in 3T3-L1 cells at day 1–2 of differentiation. 3T3-L1 preadipocyte 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 the time points as indicated in the figure. For (A,B), gene expression was measured by real-time RT-PCR and normalized to cyclophilin. For (A), n = 3–6 samples were pooled for one real-time PCR measurement. For (B), all data are expressed as mean ± SEM, n = 4. For (C), CpG methylation was measured by pyrosequencing as described in the Materials and Methods, n = 4. All data are expressed as mean ± SEM, *p < 0.05 vs. control. Con: control; AZA: 5-aza-dC.
