Figure 5.

Effect of the methylation site located upstream of the Spp1 TSS on Spp1 expression. (A) In-vitro-methylation assay of the site located upstream of the Spp1 TSS. The fragment DNA of the site located upstream of the Spp1 TSS (chr5:104846058-104846328) was enzymatically methylated in vitro by Sss1 before ligation into pGL3-Basic. The vertical axis represents the luciferase activity plus the standard deviation. (*p-value < 0.05, n = 3). (B) qPCR assay of Spp1 mRNA expression. Cyclophilin was used as the normalization control. The vertical axis represents the relative mRNA quantity plus the standard deviation (*p-value < 0.05, n = 3-4). (C) The effect of 5-dAza-C treatment on expression of Spp1 mRNA. mRNA levels of Spp1 were determined by qPCR. Gapdh was used as the normalization control. The vertical axis represents the relative mRNA quantity plus the standard deviation (*p-value < 0.05, n = 3).

Komatsu et al. BMC Medical Genomics 2012 5:5   doi:10.1186/1755-8794-5-5
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