Figure 3.

(A) T84 cells were synchronized 6 h after seeding by starvation and then stimulated to grow by adding 10% FBS in the presence of 1 μM ST or PBS. After 24 h, cells were trypsinized, pelleted, and then fixed and stained with PI. Fluorescence analysis of DNA content was performed as described in Materials and Methods, and the percentage of cells in each phase of the cell cycle is represented. Data are from a representative experiment. After synchronization by starvation (t₀), 75.7% ± 4.5 of T84 cells were in the G₀/G₁ phase of the cell cycle. (B1) T84 cells were synchronized by starvation in EMEM depleted of l-glutamine for 48 h, stimulated to proliferate with EMEM containing 10 mM l-glutamine, and incubated for the indicated time points with 1 μM ST (■) or PBS (□). [³H]Thymidine was added for the last 2 h of incubation, and the ³H incorporated into DNA was measured as described in Materials and Methods. Data reflect a representative experiment. Results from three experiments performed as described in B1 are presented in B2 as mean ± SEM. *, P < 0.05.