Figure 4

Structure/function analysis of E2F8. (A) Yeast BY1142 was transformed with the indicated E2F8 mutants (ADE2 selection) along with empty URA3 vector. Transformants were replicated onto selective plates containing 3-AT and assessed for growth after 3 days. The strength of signaling and fluorescence is reported as in Table 1. (B) Yeast cells expressing E2F8 mutants were grown in liquid culture, lysed, and examined by immunoblotting using an anti-GFP (top) antibody. The membrane was stripped and reprobed using an anti-β-actin antibody (bottom) as a loading control. The 5' UTR-E2F8-YFP construct contained the wild-type 5' UTR in addition to E2F8; the other full-length construct lacked the UTR.