FIG. 4.

G₁/S arrest does not inhibit detachment-induced Bim mRNA expression. Control and mimosine-treated MCF-10A cells were plated at 400,000 cells/well in either tissue culture-treated (attached) or Poly-HEMA-coated (suspended) six-well plates for 24 h. RNA was harvested, and double-stranded cDNA using oligo(dT) primers was synthesized from 5 μg total RNA. Real-time quantitative PCR was performed on equal amounts of total RNA as described previously (47). The relative Bim mRNA expression for each condition was plotted. Error bars represent the SEM of nine quantitative PCRs from three independent experiments.