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. Author manuscript; available in PMC: 2007 Dec 10.
Published in final edited form as: Oncogene. 2006 Dec 18;26(24):3532–3540. doi: 10.1038/sj.onc.1210143

Figure 2. E2F1 upregulation of the p57 promoter is direct.

Figure 2

A. H1299 cells were infected with the indicated adenoviral expression vectors at a MOI of 10 (based upon plaque assay), or as specifically indicated. Forty-eight hrs after infection extracts were harvested and subjected to RPA or Western. B. H1299 control cells or a derivative expressing ER-E2F1 was subjected to OHT (4-hydroxytamoxifen, 300 nM) added to medium to induce ER-E2F1 nuclear accumulation. Cycloheximide (CHX) was used at a final concentration of 10 μg/ml to inhibit new protein synthesis. Cells were harvested after 24 hrs and p57 mRNA evaluated by RPA as described above. C. Same as in B, except a Western blot was performed following OHT treatments. The lower p57 band may represent an alternatively spliced p57 mRNA (Lee et al., 1995). Cycloheximide-treated cells were not including in panel C since p57 protein could not accumulate in its presence.