Figure 1
From: Genome-wide mapping of Myc binding and gene regulation in serum-stimulated fibroblasts
Conditional deletion of the c-myc gene in quiescent cells impairs cell-cycle re-entry. (a) A schematic representation of the experimental approach. Near-confluent cultures of mycf/f;CreER fibroblasts were serum-starved for 48 h with or without OHT to induce c-myc deletion. Following stimulation with 10% fetal calf serum, RNA was isolated at the indicated time points for profiling (Figure 2). (b) c-myc mRNA levels were measured in mycf/f;CreER cells, either without (control) or with OHT to induce c-myc deletion (Δ c-myc), followed by serum stimulation, as indicated. The data were normalized to 36B4 mRNA. The average±s.d. of three independent experiments is shown. (c) Relative amount of c-myc genomic DNA measured by quantitative RT–PCR in mycf/f;CreER cells left untreated (black bars) or treated with OHT (gray bars). Values were normalized to a PCR amplicon in the Nucleolin locus, and represent the average±s.d. from three independent experiments. (d) Cumulative percentages of cells traversing S-phase over 24 h of fetal calf serum stimulation in cultures treated as above. BrdU was added to the culture medium at time 0, and incorporation was assayed by flow cytometry.
