FIGURE 3.

Trim28 forms complexes with E2F3 and E2F4. A and B, H1299 cells were transiently transfected with expression plasmids for E2F3, E2F4, or Myc-Trim28 (Myc-epitope tagged), as indicated. Whole cell lysates (WCL) of transfected cells were subjected to immunoprecipitation (IP) and then immunoblotting (IB). C, H1299 cells were transfected with expression plasmids for E2F3 or E2F4 at different amount (+: 2 μg; +++: 6 μg) or pcDNA3 (−) as indicated. WCLs of transfected cells were subjected to IP and then IB. D, A549 cells were stained (as described under “Experimental Procedures”) and examined using confocal immunofluorescence microscopy, as follows: E2F3 (red, top row), E2F4 (red, bottom row), Trim28 (green), and DAPI (blue; nuclei). Merged images are in the last column, yellow represents co-localization of E2Fs and Trim28.