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. 2005 Feb 14;168(4):553–560. doi: 10.1083/jcb.200411093

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Copyright © 2005, The Rockefeller University Press

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Figure 4. — Incomplete blockage of E2F/DP activity in HDFs expressing dn-E2F. (A and B) ChIP assays were performed using TIG-3 cells expressing dn-E2F (A), or DP1-shRNA (B) and the antibodies shown. The cyclin A2 promoter was recovered by PCR using primers flanking the E2F-binding sites in the human cyclin A2 promoter. The human β-actin promoter, which does not contain E2F-binding sites, was used as a negative control for PCR. (C) TIG-3 cells (50PDLs) expressing dn-E2F (lanes 3 and 4) or empty vector (lanes 1 and 2) were super-infected with retrovirus encoding DP1-shRNA (lanes 2 and 4) or control-shRNA (lanes 1 and 3). Total RNAs were prepared from cells at 7 d after selection with puromycin, and were subjected to RT-PCR with primers shown on the left of the figure. E2F target genes reported previously were highlighted with an asterisk. (D) Cells described in C were used in a proliferation assay performed in triplicate. Error bars indicate SD.