FIG. 10.

EID-1 interacts with p300. (A) Total protein lysates were prepared from U2OS cells, and EID-1-associated proteins were immunoprecipitated with α-EID-1 antibody. Complexes were resolved by SDS-PAGE and probed with α-p300 antibody. (B) In vitro-transcribed and -translated [³⁵S]methionine-labeled EID-1 was incubated with 1 μg of either GST or the indicated GST-p300 fusion proteins. Complexes were precipitated with glutathione-Sepharose beads and resolved by SDS-PAGE. (C) U2OS cells were transiently transfected with either 0.5 μg of GAL4-BD or GAL4–EID-1, 2 or 4 μg of CMV-p300 or CMV-p300del30, and a luciferase reporter gene under the control of multiple GAL4-binding sites. After 48 h, cell lysates were harvested and luciferase activity was assayed and normalized to that of total protein. Results are shown relative to the amount of GAL4-BD alone.