Cytoselect cell migration and invasion kit

The CytoSelect Cell Migration and Invasion kit (Cell Biolabs, Inc., San Diego, CA, USA) was used according to the manufacturer's protocol. After 24 h transfection, PCa cells were re-suspended in a serum-free RPMI-1640 medium (cat no. sh30809.01B; HyClone; GE Healthcare Life Sciences) to a density of 25×10⁴/ml. The suspended cells (200 µl) were seeded in the upper chambers, and the lower chamber contained 10% fetal bovine serum as a chemoattractant. Following incubation at 37°C for 48 h, the cells that had migrated through the membrane were stained by 0.1% crystal violet (cat no. BS234b; Biosharp, Hefei, China) at room temperature for 15 min and quantified by counting 9 independent symmetrical visual fields under a light microscope (Nikon Corporation). Data are presented as mean ± SD of three independent experiments.

The CytoSelect Cell Migration and Invasion Kit (Cell Biolabs, Inc., San Diego, CA, USA) was used according to manufacturer’s instructions. Twenty-four hours after the transfection, PCa cells were resuspended in a serum-free medium to a density of 25×10⁴/mL. The cell suspension (200 μL) was transferred to the upper chambers. The lower chamber contained 10% fetal bovine serum used as a chemoattractant. After incubation for 48 hours, cells migrated through the membrane and were fixed, stained, and quantified by counting nine independent symmetrical visual fields under a microscope. Data were presented as mean ± SD of three independent experiments.