Figure 7.

MCM7 supports the reactivated proliferation of quiescent cancer cells. (A) The reactivated cells were treated with or without higher dose of PTX (HepG2^Reac 160 nM, UMUC3^Reac 500 nM) for 3 days to induce quiescence, Western Blot were performed to examine the protein expression. (B) UMUC-3^Reac cells were treated with or without 500 nM PTX for 3 d, ChIP were done using an anti-E2F1 polyclonal rabbit antibody followed by PCR amplifying of cyclin D1 and E1 promoter regions. The ChIP results were obtained from three independent experiments, *P<0.05. (C) The reactivated cells were transfected with siRNA against MCM7 for 48 h. MCM7, Cyclin D1 and E1 proteins were detected by Western blot. (D) The reactivated cells were treated with PTX (HepG2^Reac 160 nM, UMUC3^Reac 500 nM) for 3 days to induce quiescence and then released into fresh medium, MCM7 siRNA was transfected 24h before the PTX-release, cells were collected at 0, 4, 8 and 12 h after the PTX-release, the cell proliferation was detected by EdU incorporation assay, data are shown as mean ± SD of three independent experiments, * P<0.05, and (E) MCM7 and CyclinD1 proteins levels were detected by WB. GAPDH was used as loading control.