A, indicated model systems were cultured in media containing complete serum, harvested, and lysed; total protein was separated by SDS-PAGE, transferred.

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A, indicated model systems were cultured in media containing complete serum, harvested, and lysed; total protein was separated by SDS-PAGE, transferred to PVDF, and immunoblotted for indicated proteins. A, indicated model systems were cultured in media containing complete serum, harvested, and lysed; total protein was separated by SDS-PAGE, transferred to PVDF, and immunoblotted for indicated proteins. Representative image of at least 3 independent experiments is shown. B, Poly(ADP-ribose) (PAR) ELISA was conducted on whole-cell lysates of indicated model systems. Data reflect averages and SD of at least 3 independent experiments, each conducted with technical and biologic triplicates. C, top, indicated model systems were steroid deprived for 72 hours, then treated for 30 minutes with Poly(ADP-ribose) vehicle or ABT888 (2.5 mmol/L). Cells were harvested and qPCR analyses were conducted for indicated target genes. Data reflect averages and SE of at least 3 independent experiments, each conducted with biologic triplicates. Results were normalized to 18S and control is set to “1.” Bottom, indicated model systems were treated as above, then harvested, and lysed; total protein was separated by SDS-PAGE, transferred to PVDF, and immunoblotted for indicated proteins. Representative image of at least 3 independent experiments is shown. D, C4-2 cells were steroid deprived for 72 hours, then treated for 30 minutes with vehicle or ABT888 (2.5 mmol/L). Cells were fixed and lysed, and ChIP for AR and PARP-1 was conducted. DNA was isolated from IPs and used in qPCR reactions for indicated genomic loci. Data are representative of at least 3 independent experiments, each conducted with technical triplicates, and depicted as the average of immunoprecipitated signal to input signal and SE. 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