Figure 7
gcCITED3a and gcCITED3b physically interact with CBP/p300. A. Immunoprecipitation and Western blot analysis. CHO cells were separately transfected with pGFP-gcCITED3a, pGFP-gcCITED3aΔCR2, pGFP-CITED3b, or pGFP-CITED3bΔCR2, and whole cell lysates prepared and immunoprecipitated with anti-CBP antibody (Santa Cruz). The immunoprecipitates were resolved on 12% denaturing SDS-PAGE and Western blot analysis was carried out to detect GFP fusion proteins using anti-GFP antibody (Santa Cruz). B. GST pull-down assays. GST protein only or GST protein fused to gc-p300/CH1 (CH1 domain of grass carp p300) was purified from bacterial culture and immobilized on glutathione-Sepharose beads. In-vitro-transcribed and -translated [35S]-methionine-labeled gcCITED3 or gcCITED3ΔCR2 was incubated with purified GST-fused CH1 or GST alone as indicated in the figure.