Evaluating VIRMA Knockdown in PC-3 Cells

To evaluate the impact of VIRMA knockdown in PC-3, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT; Sigma-Aldrich, St. Louis, MO, USA) assay was performed. To determine differences in cell proliferation the Cell Proliferation ELISA BrdU assay (Roche Applied Sciences, Basel, Switzerland) was used. Invasion ability of cells was analyzed using BD Biocoat™ Matrigel Invasion Chambers (BD Biosciences, San Jose, CA, USA). The procedures followed the published report [23 (link)]. For wound-healing assay cells were seeded and grown to 100% confluence and then scratched with a sterile 200 μL pipette tip to create an artificial wound. At 0, 4, 8, and 12 h after wounding, phase-contrast images of the wound healing process were photographed with a 10x objective lens. Eight images per condition were analyzed to determine averaging position of the migrating cells at the wound edges.