Figure 1

Overexpression of RASSF1A delays G₁–S progression in a p53-dependent manner. (A) HCT116 cells (p53^+/+, p53^-/- and p53^-/- complemented with HA–p53) were transfected with pcDNA-CD4 and HA–RASSF1A. The cell-cycle profile of CD4-positive cells was determined 36 h after transfection. Data are expressed as meansstandard errors of values from three independent experiments. (B) Cells in (A) were analysed by immunoblotting for the indicated proteins. Asterisk indicates nonspecific bands. (C) U2OS cells incubated for the indicated times after transfection with HA–RASSF1A were exposed during the final 1 h to 10 M BrdU and then analysed by flow cytometry to determine the percentage of BrdU-positive cells. Data are expressed as meansstandard errors of values from three independent experiments. (D) U2OS cells were co-transfected with pGL3-p53-responsive reporter and pcDNA3--Gal reporter constructs and the indicated combinations of pcDNA3-p53 (wt), pcDNA3-p53-R248W (mt) and HA–RASSF1A. Cell lysates were subjected to immunoblot and luciferase assays. Luciferase activity was normalized to -galactosidase activity and expressed relative to that in cells transfected with an empty vector. Data are expressed as meansstandard errors of values from three independent experiments.