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. 1997 Jul 8;94(14):7245–7250. doi: 10.1073/pnas.94.14.7245

Figure 2.

Figure 2

Specificity in the activation of target genes by E2F family members. (A) Activation of DNA synthesis and cell cycle regulatory genes by E2F proteins. REF52 cells were deprived of serum for 48 hr and then infected with the indicated recombinant viruses as described in Fig. 1B. Con+ was stimulated by the addition of 10% serum, and the others remained in 0.25% serum. The cells were harvested at either 18 hr (Con+) or 21 hr (others) postinfection, and poly(A)+ RNA was isolated. Poly(A)+ RNA (derived from 330 μg of total RNA per lane) was then separated by agarose gel electrophoresis, transferred to nylon membrane, and probed with the indicated cDNAs. Equivalent loading of RNA was confirmed by probing with the GAPDH cDNA. The hybridizing species in the thymidine kinase blot unique to the E2F4 sample represents cross-hybridization with a viral specific RNA. (B) Induction of CKI genes. REF52 cells were infected and harvested as described in A. Poly(A)+ RNA (derived from 400 μg of total RNA per lane) was then separated by agarose gel electrophoresis, transferred to nylon membrane, and probed with the indicated cDNAs. Exon I-specific fragments were used as probes for p15INK4B, p16INK4A, and p19ARF.