FIG. 4.
EID-1 associates with Rb in mammalian cells. (A) In vitro-transcribed and -translated [35S]methionine-labeled EID-1 or EID-1(C180G) was incubated with 1 μg of either GST or GST-Rb. Complexes were precipitated with glutathione-Sepharose beads and resolved by SDS-PAGE. (B) L17 cells were transiently transfected with FLAG epitope-tagged EID-1 or EID-1(C180G). Cell lysates were immunoprecipitated with anti-Rb antibody, immunoprecipitates were resolved by SDS-PAGE and analyzed by immunoblotting with anti-FLAG antibody. Total cell lysates were immunoblotted with anti-FLAG antibody to ensure the equivalent expression of proteins.