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. 2008 Oct 1;3(10):e3306. doi: 10.1371/journal.pone.0003306

Figure 3. Genomic DNA methylation status of reactivated genes in leaf and Arabidopsis cell suspensions (ACS).

Figure 3

(A) Bisulfite genomic sequencing of twelve clones of the TTG1 (three different regions), GSTF5, SUVH8, Fimbrin and CCD7 promoters. In the schematic representations of the methylation status of each CpG dinucleotides black and white dots indicate methylated and unmethylated CpGs, respectively. ACS IMP: Arabidopsis cell suspensions at intermediate passage. (B) Expression profiles of reactivated genes in leaves and Arabidopsis cell suspensions determined by quantitative RT-PCR assays. TTG1, GSTF5, SUVH8, Fimbrin and CCD7 are more strongly expressed in leaf tissues. Expression in cell suspensions can be restored by treatments with the demethylating drug ADC. ACTIN was used as a control. (*) the expression level of TTG1 in leafs is 80-fold higher than in ACS. (**) the expression level of GSTF5 and CCD7 in leafs is 40-fold higher than in ACS. (C) Promoter CpG island hypermethylation is associated with changes of histone modifications. Chromatin immunoprecipitation analysis of the histone-modification status of the promoters of the TTG1, GSTF5, and SUVH8 genes. NAB is the control without antibody. The promoter region of ACTIN is used as a control. AcH3, acetylated histone H3; AcH4, acetylated histone H4; 3mK4, trimethyl-lysine 3 histone H3.