Skip to main content
. Author manuscript; available in PMC: 2012 Dec 8.
Published in final edited form as: Cancer Lett. 2011 Jul 22;311(2):177–186. doi: 10.1016/j.canlet.2011.07.015

Fig. 3.

Fig. 3

Effect of cPAcP expression on VPA growth suppression and VPA effect on androgen sensitivity of PCa cell lines. (A) LNCaP C-81 cells were plated at a density of 1×105 cells/well in 6-well plates for 72 hr and then transfected with PAcP shRNA-126 plasmids. Control cells were transfected with the vector containing the scramble DNA. Five hours after transfection, the cells were fed with RPMI medium containing 10% FBS for 24 hr. The cells were then treated with 1mM VPA for 48 hr. The cell numbers were counted. The ratio of cell growth was calculated by normalizing the cell number to that of the control cells transfected with the control vector and treated with the solvent alone. The result shown is the average from two sets of independent experiments in triplicates (left panel, n= 3×2). Total cell lysate proteins from 3-day VPA treatments were analyzed for cPAcP protein expression. β-Actin was used as a loading control (right panel). (B) LNCaP C-81; (C) C4-2; (D) MDA PCa2b-AI cells were plated at a density of 3×104, 3×104 and 1×105 cells/well, respectively, in 6-well plates for 72 hr and then treated with 1 mM VPA. Control cells were treated with solvent alone. After 48 hr VPA treatment, cells were maintained in a steroid-reduced medium in the absence or presence of 10 nM DHT for 48 hr. The cell numbers were counted. The ratio of cell growth was calculated by normalizing the cell number to that of the control cells without any treatment (column #1). The result shown is the average from three sets of independent experiments in triplicates (n= 3×3). Total cell lysate proteins from 3-day DHT treatment were analyzed for cPAcP, PSA, AR protein expression. The AR protein in Fig. 3B was obtained after 3-hr hybridization with anti-AR Ab followed by an exposure time period of less than 1 min; while the AR proteins in Fig. 3C & 3D were obtained after overnight hybridization with anti-AR Ab and with an exposure time period of 1 hr. β-Actin was used as a loading control and it was obtained with the same reaction time periods.