Figure 11 .

Two GADR proteins, GADR-5/Y71A12B.17 and GADR-6/F47G4.2, physically interact with both CUL-2 and ELC-1 when coexpressed in human cells. FLAG-tagged GADR-5, GADR-6, and ZYG-11 were coexpressed in HEK293T cells with CUL-2–Myc or HA–ELC-1 as noted by (+) symbols above the lanes. Anti-FLAG immunoprecipitations (IP) and lysates were analyzed by Western blot using anti-FLAG, anti-HA, or anti–CUL-2 antibodies. A cross-reacting band serves as a loading control. Note that both GADR-5 and GADR-6 bind CUL-2 and ELC-1 analogous to the known substrate recognition subunit ZYG-11. The smearing and additional lower bands for FLAG–GADR-5 presumably arise from partial degradation of the protein in HEK293T cells. * denotes the heavy chain of IgG used in the IP; ** marks nonspecific band (which comigrates with lower band of CUL-2 in the first four samples).