Fig. 3.

Effect of p-DGA and m-DGA on apoptosis and AR in human PCA cells. Human PCA 22Rv1 and LNCaP cells were treated with vehicle or m-DGA or p-DGA (50 and 100 µg/mL) for 24 and 48h. (A) After 48h of treatment, both adherent and non-adherent cells were collected, stained with annexinV/PI and analyzed by flow cytometry for the apoptotic cell population. The data are presented as mean (n=3) ± SEM and represents two independent experiments. *, P<0.001; $, P<0.05 (B) Whole cell lysate were prepared after treating 22Rv1 and LNCaP cells with m-DGA or p-DGA for 48h and used to analyze the protein expression of cleaved caspase 3 (CC3), cleaved caspase 9 (CC9), and cleaved PARP (Cl. PARP) by western blotting. (C) Western blotting was performed for AR and PSA; and membranes were re-probed with β-actin to check equal protein loading. For the secreted PSA expression, media was collected and analyzed for PSA expression by immunoblotting. In each case, the media loading volume was normalized with the respective protein value of the cell lysate. The densitometry data presented below the bands are “fold change” as compared to control after normalization with respective loading control (β-actin). ND: Not detectible.