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. 2006 Mar 1;20(5):613–623. doi: 10.1101/gad.1345006

Figure 4.

Figure 4.

The marked box region of E2F1 is required for Jab1 binding and enhancement of apoptosis. (A) Diagram of E2F chimeras used in these studies. The chimera 111331 is composed of the E2F1 cyclin A-binding, DNA-binding, DP1 dimerization, and transactivation/Rb-binding domains; however, the marked box and adjacent region originate from E2F3. The reverse chimera, 333113, is E2F3 with the E2F1 marked box and adjacent regions replacing the homologous sequences from E2F3. The chimeras 111311 and 333133 substitute only the marked box region between the genes. (B) HEK293 cells were transfected with HA-E2Fs and with pcDNA3 or pcDNA3–5×-Myc-Jab1. Myc-Jab1 complexes were immunoprecipitated with anti-Myc antisera (9E10) and protein G beads, separated by SDS-PAGE, and immunoblotted with anti-HA antisera. (C) Quiescent REF52 cells were infected with AdCMV, AdE2F1, AdE2F3, or adenovirus expressing chimeric E2Fs along with either control adenovirus or AdJab1. Cells were harvested at 40 h post-infection and processed for active caspase-3 levels by FACS analysis.