Fig. 7

Differential interaction of MCI and GMNC with E2F4 and E2F5. (A) Co-immunoprecipitation experiments using mouse (Mm, Mus musculus) proteins show that MCI effectively interacted with both E2F4 and E2F5 in the presence of DP1 (lane 1, IP panel, anti-Myc blot); very little E2F4 co-precipitated with GMNC and DP1 (lane 5, IP panel, anti-Myc blot). The weak presence of E2F4 in the GMNC IP is marked by the black asterisk. (B) The C-terminal domain of MCI accounts for effective interaction with both E2F4 and E2F5. GMNC showed minimal interaction with E2F4 (lane 1, IP panel, anti-Myc blot). In comparison, the GM (engineered by replacing the C-terminal domain of GMNC with that of MCI) chimera could co-precipitate with both E2F4 and E2F5, in the presence of DP1 (lane 5 and 7, IP panel, anti-Myc blot). The black asterisk marks the E2F4 band that is absent in lane 1 (IP panel, anti-Myc blot). Human (Hs, Homo sapiens) proteins were used for this experiment. The E2F and DP1 proteins were tagged N terminally with the Myc epitope, and the GMNC and MCI proteins were tagged N terminally with the HA epitope. TCL, total cell lysate; IP, immunoprecipitation. Data are representative of two independent biological replicates.