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

Figure 2.

Figure 2.

Depletion of DP1 causes reduction of E2F target gene expression in HDFs. (A) 20 μg of total RNA prepared from TIG-3 (50PDLs), HeLa, U2OS, HT29, and 293T cells transfected with or without DP2 expression plasmid were subjected to Northern blot analysis using the same activity of indicated radiolabeled probes. (B and C) Early passage TIG-3 cells (45PDLs) were infected with retrovirus encoding DP1-shRNA (lane 2) or control-shRNA (lane 1). Cell extracts were prepared from cells at 7 d after selection with puromycin, and were subjected to RT-PCR (B) or Western blotting (C) with primers or antibodies shown at left. E2F target genes reported previously were highlighted with an asterisk. CDK4 was used here as a loading control. (D) EMSA was performed using extracts from cells infected with retrovirus encoding control-shRNA (lanes 1 to 5) or DP1-shRNA (lanes 6 to 10) in the absence or presence of antibodies shown at top. The DNA-binding activity, which is resistant to any available E2F antibodies, was marked by an asterisk.