Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Figure 1. AtMBD-GFP proteins display differential subnuclear localization. (A) AtMBD6 is accumulated at chromocenters showing preference for two perinucleolar domains. Sequential confocal optical sections (1-9, slices of 0.3 μm) of Arabidopsis nucleus transiently expressing AtMBD6-GFP. Note the strong GFP signals adjacent to the nucleolus (nuc, arrowheads in slice 6) and the weaker signals at the periphery (arrows) representing chromocenters. (B) AtMBD2 is localized to euchromatic regions in the Arabidopsis nucleus. Confocal optical sections (1-6) of Arabidopsis nucleus transiently expressing AtMBD2-GFP. Numbers at the bottom of each slice indicate the distance in μm from reference slice No. 1 (0.0). Note the exclusion of the GFP signal from chromocenters (arrowheads in slice 3) and the nucleolus (nuc).
• Supplemental Figure 2 - Figure 2. AtMBD2-GFP is tightly bound to chromatin. Nuclei prepared from leaves of transgenic Arabidopsis expressing AtMBD2-GFP were extracted with increasing concentrations of NaCl. Soluble (S) and insoluble pellet (P) fractions were analyzed for the presence of AtMBD2-GFP or histone H3 methylated at lysine 4 by immunoblotting, using anti-GFP (αGFP) and anti-dimethylated K4 histone H3 (αK4m2H3), respectively.
• Supplemental Figure 3 - Figure 3. Localization of AtMBD6-GFP to chromocenters in wild-type nuclei is disrupted in ddm1-2, met1-1,and met1-3. (A) Randomly selected nuclei expressing AtMBD6-GFP are shown. Note that subnuclear distribution of AtMBD6-GFP is strongly affected in ddm1-2 compared to met1-1 and met1-3 mutants. (B) Mean coefficient of variation and the standard error of mean (SEM). Coefficient of variation was determined from the frequency distribution of the pixel intensity within the nucleus. Six nuclei were examined in each population yielding the following coefficient of variation: Wild type, 0.772 ± 0.18; ddm1-2, 0.114 ± 0.029; met1-1, 0.239 ± 0.06; met1-3, 0.225 ± 0.062. Statistical significance between ddm1-2 and the other lines was determined by T test. Note that differences between lines are statistically significant.