Harvested xenografts were bisected through the longest dimension, fixed in 10% neutral-buffered formalin, and embedded in paraffin. Preparation of formalin-fixed, paraffin-embedded tissue sections, routine hematoxylin and eosin (H&E) staining, and IHC were conducted as previously described (20 (link)). Residual tumor volumes in dormant prostate cancer (12 weeks postcastration time point) were determined by microscopic imaging analyses as previously described (20 (link)).
For IHC, a rabbit monoclonal anti—androgen receptor (AR) antibody (1:100, Abcam), a rabbit monoclonal anti-PSA antibody (1:200, Abcam), a monoclonal mouse anti-human Ki-67 antibody (1:25, Dako), and a rabbit monoclonal anti—caspase-3 antibody (1:50, Cell Signaling Technology) were used as primary antibodies. For quantification of Ki-67 or caspase-3 IHC stains, positive cells per 5,000 cancer cells were counted by two independent investigators.
For IHC, a rabbit monoclonal anti—androgen receptor (AR) antibody (1:100, Abcam), a rabbit monoclonal anti-PSA antibody (1:200, Abcam), a monoclonal mouse anti-human Ki-67 antibody (1:25, Dako), and a rabbit monoclonal anti—caspase-3 antibody (1:50, Cell Signaling Technology) were used as primary antibodies. For quantification of Ki-67 or caspase-3 IHC stains, positive cells per 5,000 cancer cells were counted by two independent investigators.