FIGURE 2.

Pygo2 interacts with DDB1. A, identification of DDB1 and VPRBP as candidate Pygo2-interacting proteins. A GST pull-down assay was performed as described under “Experimental Procedures.” Left, silver staining showing the banding patterns of GST versus GST-NHD complexes. Right, unique peptides identified for DDB1 and VPRBP. B–D, physical association between Pygo2 and DDB1 in HEK293T (B and C) or HC11 (D) cells. For B and C, HEK293T cells were transfected with FLAG-Pygo2 and GFP-DDB1 as indicated. Twenty-four hours later, whole cell lysates were immunoprecipitated (IP) using FLAG M2 beads (B) or anti-GFP antibody (C) and then immunoblotted using the indicated antibodies. D, whole cell lysates from HC11 cells were subjected to immunoprecipitation with anti-DDB1 antibody or IgG as indicated. The immunocomplexes were probed separately for Pygo2 and DDB1.