Fig. 1
E2F3c and E2F3d are newly identified E2F3 products. a–f HFFs were infected with control (Vec) or E1A-expressing retroviruses. a Cell lysates were analyzed by immunoblotting to examine the protein levels of E2F target genes. b Cell lysates were immunoprecipitated with an antibody against the C-terminal domain of human E2F3 proteins and probed with an antibody against the N-terminal domain of human E2F3a protein. c Total mRNAs were subjected to RT-PCR. The RT-PCR products observed in E1A-expressing cells (bands 1–3) were extracted separately and further amplified in the second reaction using inner primers nested within the first primers. d mRNA structure of newly identified E2F3 members. Schematic diagrams represent the exon composition of the RT-PCR products. e Domain structure of E2F3 members. f cDNA samples were subjected to qRT-PCR using primer sets specific for each E2F3 member. Values shown represent the means of three independent experiments. Error bars represent SD. **P < 0.01. g cDNA samples from asynchronously growing (untreated) and quiescent (serum starved) HFFs were subjected to qRT-PCR as described in f. Values shown represent the means of three independent experiments. Error bars represent SD. *P < 0.05 and **P < 0.01. h 293 T cells were transfected with expression vectors encoding Flag-tagged individual E2F3 members. Cell lysates were subjected to immunoprecipitation with anti-Flag antibody-agarose beads followed by immunoblotting with antibodies directed against Flag or the N- or C-terminal domain of human E2F3a protein. Full-size scans of immunoblots are shown in Supplementary Information
