24 well 8 μm pores transwell plates

24-well (8 μm pores) transwell plates (Corning, Lowell, MA) were used for invasion assays. For in vitro invasion assays, the upper chambers of the transwells (8 μm) were pre-coated with diluted matrigel (BD Biosciences, Sparks, MD). Before performing invasion assays, PCa cells were cultured with either mast cells or CM for 48 hrs, and then wash out mast cells with PBS 3 times and then collected the co-cultured cells for further experiments. In brief, 10⁵ PCa cells (in serum free media) and 10% serum containing media were plated in the upper chambers. After 48 hrs incubation, invaded cells were stained with 0.1% crystal violet, and positively stained cells were counted. The cell numbers were calculated from counting six random fields. Quantitation indicates mean of triplicate repeats ± SEM.

24-well (8 μm pores) transwell plates (Corning, Lowell, MA) were used for invasion assay. For in vitro invasion assays, the upper chambers of the transwells (8 μm) were precoated with diluted matrigel (BD Biosciences, Sparks, MD). Before performing invasion assays, PCa cells were cocultured with pre-adipocytes for 48 hrs and then trypsinized. 10⁵ PCa cells (in serum free media) and media containing 10% serum were plated in the upper and lower chambers, respectively. After 48 hrs incubation, invaded cells were stained with 0.1% crystal violet, and positively stained cells were counted. The cell numbers were calculated from counting six random fields. Quantitation indicates mean ± SEM of triplicate repeats.