Figure 4

Validation of the methylation site located upstream of the Spp1 TSS. (A) Epigenetic features of the methylation site located upstream of the Spp1 TSS assessed using the UCSC Genome browser. The Spp1 gene body is shown in dark blue. The methylation site upstream of the Spp1 TSS is shown as a red rectangle on the chromosome diagram and on the custom track. ChIP-seq signals for H3K4 me1 and H3K4 me4 in the liver (orange), for RNAP II in the liver and p300 in MEL leukemia (light blue), and for the DNase I hotspot in the liver (green) are represented as density plots. (B) MBP-IP assay of the methylation status of the site located upstream of the Spp1 TSS. The CCl₄ sample was normalized to the control sample. The vertical axis represents fold changes plus the standard deviation (**p-value < 0.01, n = 3-4). (C) Bisulfite modification assay of the methylation status of the site located upstream of the Spp1 TSS. The CCl₄ sample was normalized to the control sample. The vertical axis represents percentage of methylated sites in the site located upstream of the Spp1 TSS (n = 10).