Effect of LY 294002 on the induction of cyclin D1 and c-myc mRNA. MCF-7 cells were starved of serum and E2, in the presence or absence of ICI 182780, for 48 h. In one series of dishes, LY294002 was added for the last 3 h of serum deprivation (time −3 h to 0 h). The cells were then stimulated with E2 (1 μM) or insulin (1 μM) for 4 h. The cells were then harvested for the isolation of RNA and RT-QPCR analysis. Fold induction of ΔΔCT (means of three independent experiments performed in triplicate) related to the values at t = 0 in cells starved of serum in the absence of ICI 182780, are presented. The differences between bars 3 vs 11, 4 vs 12, 6 vs 8 and 9 vs 11 are significant at p ≤ 0.05 (*) or p ≤ 0.01 **).