Matrigel transwell invasion assay

The ability of DU145 and PC3 cells to invade through Matrigel was examined using Matrigel coated invasion chambers (#354480, Corning) according to the manufacturer’s instructions. Samples were prepared in triplicates, 1 × 10⁵ cells were seeded into the top chamber of 24-well insert in 0.5 ml 0.1 % BSA-containing RPMI SF media. Medium with 10 % FCS was used as a chemoattractant in the bottom chamber. After 24 hours, residual cells on the top surface of insert were removed, membranes were fixed with cold methanol and nuclei were stained with DAPI. Nuclei were imaged from 9 fields of each membrane, and counted with ImageJ. Average nuclei count for each cell type was obtained, normality of the data was tested with the Shapiro-Wilk test and statistical significances were obtained by comparing the average nuclei counts with one-way ANOVA test and pairwise comparisons were made with the Tukey HSD post hoc test.