Figure 5.

LEDGF depletion induced S-phase and G2–M cell cycle arrest and failure to accumulate cyclin B1. (a) Si-Control and Si-LEDGF stably transfected DU145 cells were plated in 100 mm dishes and cultured for 24 h. Cells were harvested and hypotonically lysed in a PI solution to stain the DNA. Nuclei were analyzed for DNA content by flow cytometry. Percentage of cell population at G1, S and G2 phases are shown on the right (compare Si-Control and Si-LEDGF). (b) Western analysis to examine altered expression of cell cycle regulatory proteins. Decreased expression of cyclin B1 was observed in LEDGF-depleted cells (upper panel, lane 2). Lower panel, membrane probed with β-actin antibody as loading/internal control. Each band of blot was quantified using densitometer shown on the right (black bar versus gray bar, **P<0.001)