Figure 3

Regulation of Ets1, Rrm2 and Ntf3 expression by Rb-E2F complex. (A) Diagram showing a dominant negative (DN) form of E2F where the E2F DNA binding domain is fused to the mutant hydroxyl-tamoxifen (OHT)-dependent estrogen receptor (m-ER) displacing E2F and Rb-E2F from a promoter following OHT treatment. IPTG-inducible p16 expression is shown leading to assembly of E2F-Rb complexes at target genes [40]. (B) Human U2OS cells were treated with IPTG, OHT or IPTG + OHT as described in Material and Methods. Real time PCR was used to measure mRNA levels of Ets1 (C), Rrm2 (D) and Ntf3 (E). “Ctrl” indicates untreated control cells. Results were normalized to β-actin mRNA. Rrm2, ribonucleotide reductase, Ntf3, neurotrophin 3.