Figure 4. PC3 cells undergo growth arrest upon treatment with (S)-2.

(A) – Cells (10⁵) were seeded in 6-well plates and allowed to attach overnight. The day after (S)-2 was added at the indicated concentrations and cell number was determined along the next three days. (B) – Cells were incubated for 24 h with 2.5 µM (S)-2 to determine the % of PI-stained cells in different phases of the cell cycle as determined by flow cytometry. Pictures of either untreated and treated cultures were taken with the aid of a phase-contrast microscopy. (C) – p21 mRNA levels in PC3 cells from cultures treated without/with (S)-2 were measured by quantitative real-time PCR. (D) – Comparative Western blot analysis of acetyl-H3 levels in cell extracts from PC3 treated with either (S)-2 or SAHA; α-tubulin was used as loading control.