The chicken homolog of MBD2 is a bona fide methyl-CpG-binding protein and is a component of the MeCPC. (A) DNA sequences of the 50-bp probes used for EMSA with cMBD2. The sequence is derived from the first exon of the ρ-globin gene. The unmodified sequence contains 1 CpG (a1CpG). The sequence was modified to contain no CpGs (a0CpG), 2 CpGs (a2CpG), and 3 CpGs (a3CpG). All CpGs are indicated in bold. Bases that have been mutated compared with the wild-type ρ-globin gene are underlined. (B) cMBD2 is a bona fide MCBP in vitro. The probes a0CpG, a1CpG, a2CpG, and a3CpG were methylated and incubated with recombinant cMBD2. The binding reactions were subjected to EMSA. cMBD2 forms a complex (indicated by the arrow) with the probes containing methyl-CpGs. (C) Anti-cMBD2 antibodies supershift the MeCPC. Inclusion of V2 anti-cMBD2 IgG, but not control IgG, retarded the mobility of the MeCPC complex (supershift) during EMSA.