FIGURE 2

Cebpd-deficient 3T3 fibroblasts display reduced serum dependence and contact inhibition. 'WT' and 'KO' data represent the means.d. of the WT#1 and WT#2, and KO#1 and KO#2 lines, respectively. (a) Cell death upon serum withdrawal. Cells were seeded at 3 10⁵ cells per 100 mm dish. The next day, the medium was changed to 0 or 0.07% serum. After 2 days, FACS analysis of the DNA content was performed. The sub-G1 populations are shown as a measure of cell death. Results represent the means and s.d. of duplicate experiments. (b) Cell survival upon serum withdrawal. Cells were seeded with 10% serum in 24-well plates at 2 10⁴ cells per well. The following day, three wells were used to determine the cell number. For the remaining wells, the medium was changed to 0 or 0.07% serum following two washes with PBS. The number of cells per well was determined 48 h later. The data are expressed as relative cell survival (cell number at the time of serum reduction=1). A representative experiment performed with triplicate wells for each cell line is shown. (c) Cell cycle distribution under subconfluent and confluent culture conditions. The cells were seeded at 3 10⁵ cells per 100 mm dish, collected on day 2 (G, growing) or day 4 (C, confluent), and FACS analysis of the DNA content was performed to determine the cell cycle distribution (means.d. of % cells in G0/1 and S+G2/M of three independent experiments)