Figure 4.
Reactivation of the Silenced Pro35S:NPTII Gene Is Independent of DNA Methylation but Related to Histone Modification.
(A) DNA gel blot analysis of genomic DNA digested with methylation-sensitive enzymes DdeI (CTNAG) and XmiI (GTMKAC) hybridized with the 35S promoter as probe.
(B) Bisulfite sequencing analysis of cytosine methylation in the upper strand of the 35S promoter (from 61 to 400, GenBank accession number AJ007625).
(C) ChIP analysis of histone H3 acetylation and H3K9 dimethylation in 35S promoter region. Chromatin was precipitated using specific antibodies against acetylated histone H3 and methylated H3 (anti-H3K9). Equal input used for ChIP was confirmed by equal PCR amplification. Actin was used as a control for anti-AcH3 and anti-H3K9 precipitation. NoAb, precipitation without antibody as a negative control.
(D) The expression of Ta3 in ror1 ros1, ros1, ddm1, and wild type by RT-PCR. Tubulin was used as a control.