A, left: LNCaP cells were cultured in media containing complete serum, treated for 24 hours with either ethanol control (0.01%), ABT888 (2.5 mmol/L), or.

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A, left: LNCaP cells were cultured in media containing complete serum, treated for 24 hours with either ethanol control (0.01%), ABT888 (2.5 mmol/L), or CSDX (1 mmol/L). A, left: LNCaP cells were cultured in media containing complete serum, treated for 24 hours with either ethanol control (0.01%), ABT888 (2.5 mmol/L), or CSDX (1 mmol/L). Cells were harvested, and quantitative PCR (qPCR) analyses for indicated target genes were conducted. Data reflect averages and SE of at least 3 independent experiments, each conducted with technical triplicates. Results were normalized to 18S and control is set to “1.” Middle, LNCaP cells were cultured, treated, and harvested as in A, then lysed; and total protein was separated by SDS-PAGE, transferred to polyvinylidene difluoride (PVDF), and immunoblotted for indicated proteins. Representative image of at least 3 independent experiments is shown. Right, same as left, except cell model is VCaP. B, LNCaP cells were steroid deprived for 72 hours, then pretreated for 30 minutes with either vehicle or ABT888 (2.5 mmol/L), stimulated with DHT (1 nmol/L, 24 hours) or ethanol control. Cells were harvested and qPCR analyses for KLK3/PSA were conducted. Data reflect averages and SE of at least three independent experiments, each conducted with technical triplicates. Results were normalized to 18S and control is set to “1.” C, LNCaP cells were cultured in media containing complete serum, treated for 24 hours with either ethanol control (0.01%), ABT888 (2.5 mmol/L), CSDX (1 mmol/L), or a combination of ABT888 (1.75 mmol/L) and CSDX (0.5 mmol/L). Cells were harvested, and qPCR analyses for indicated target genes were conducted. Data reflect averages and SE of at least 3 independent experiments, each conducted with technical triplicates. Results were normalized to 18S and control is set to “1.” Statistical significance was determined using Student t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Matthew J. Schiewer et al. Cancer Discovery 2012;2:1134-1149 ©2012 by American Association for Cancer Research