Comparison of rare vs macroscopic human prostate cancer cells in mouse bone

Luciferase and GFP labeled Du145 prostate cancer cells were injected into the left cardiac ventricle of CB17 SCID mice and allowed to grow until tumors were detectable in bones. FACS was used to isolate prostate cancer cells as described (Cackowski et al, J Cellular Biochemistry 2017) from bones with detectable tumor or the remaining pelvis, lumbar vertebrae, femorae, tibiae and humorae which had tumor cells present but below the limit of detection by imaging. After RNA isolation and cDNA synthesis, the samples were profiled by bulk RNA sequencing. Pairwise comparison of bones grossly involved with tumor vs. bones where tumor was not detected by imaging, from two mice.