Figure 3

PCA3 knockdown induces cell cycle arrest and apoptosis in LNCaP cells. (A) Representative histograms of LNCaP cells 36 h after transfection with siPCA3 or siScr (left panels). Flow cytometry was used to quantify the percentage of cells undergoing apoptosis (cells in sub G0-G1), which are shown in the bar graph on the right panel. The data represent the means ± S.D. from three independent experiments (right panel). The asterisks indicate significant differences between siPCA3 and siScr control-treated groups (*** p < 0.001). (B) Nuclear morphological changes characteristic of apoptotic cells in LNCaP cell cultures transfected with siPCA3 or siScr (left panels) were analyzed. A representative experiment is shown. Following fixation and staining with DAPI, LNCaP cells transfected with siPCA3 or siScr were examined and photographed using a fluorescence microscope as described in the Methods section. The number of pyknotic nuclei in each experimental sample was counted, and is represented on the bar graph in the right panel as the percentage of cells with pyknotic nuclei in relation to the total number of cells counted. (C) Nuclear morphological changes characteristic of apoptotic cells in PC3 cell cultures transfected with siPCA3 or siScr (left panels) were analyzed, and the results represented as described for LNCaP cells in (B). The data represent the means ± S.D. from three independent experiments (right panel).