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. 2006 Aug 7;34(13):e96. doi: 10.1093/nar/gkl527

Figure 5.

Figure 5

Wild-type NxMBD proteins localize to CpG-rich heterochromatin in a DNA methylation dependent manner in vivo and can deliver a functional domain to a methylated reporter gene. The indicated NxMBD proteins were expressed in wild-type (A) or DNA methylation deficient Dnmt1-null (B) mouse fibroblasts that were fixed and stained using an anti-HA antibody. Left panels show anti-HA signals and right panels show DNA counterstained with DAPI. (A) Co-localization of wild-type 1xMBD, 4xMBD and the 4xMBD–VP16 fusion proteins with heterochromatic foci in interphase cells. (B) Dispersed nuclear localization of 1xMBD and 4xMBD in Dnmt1-null cells that lack DNA methylation. (C) The 4xMBD–VP16 fusion protein specifically activates a methylated reporter construct in vivo. Non-methylated or methylated reporter constructs were co-transfected alone, or with 4xMBD (500 ng), 4xMBD–R22A (500 ng) or a 4xMBD–VP16 fusion construct (250 ng). The average relative reporter activity for a triplicate experiment is shown. The error bars represent ±SD.