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. 2009 May 29;18(17):3178–3193. doi: 10.1093/hmg/ddp256

Figure 2.

Figure 2.

DNMT3B co-immunoprecipitates (co-IPs) with CENP-C and CENP-A in mammalian cells. (A) Ectopically expressed DNMT3B and CENP-C interact in mammalian cells. The indicated constructs were transiently transfected into 293T cells then whole cell extract was prepared. The immunoprecipitating antibody (IP Ab) is indicated along the top of the western panel and the antibody used in western blotting (WB) is shown below it. Reciprocal co-IPs were performed. (B) Endogenous DNMT3B and CENP-C co-immunoprecipitate from HeLa nuclear extract (M phase). In the left panel, two different CENP-C antibodies (labeled 1 and 2) are used in the co-IP. In the right panel, the positive control of CENP-C interacting with CENP-A is also shown. (C) Confirmation that DNMT3B is a centromere-associated protein as shown by its ability to co-IP with the centromere-specific H3 variant CENP-A. The indicated constructs were co-transfected into 293T cells and used for reciprocal co-IP. Input—whole cell (A and C) or nuclear extract (B) prior to co-IP, IgG—negative control co-IP with species matched normal IgG.