Fig. 7. Ral-coupled Exocyst activity is required for invasive motility of prostate tumor cells. (A) Wound healing assay. Confluent monolayers of R3327-5'A cells, transfected with siRNAs specific for Sec5, Sec6 or a control non-targeting siRNA, were experimentally wounded by scratching, and analysis was performed as described in the Materials and Methods. (B) Matrigel invasion assay. Non-metastatic R3327-5'B cells or metastatic R3327-5'A cells transfected with indicated siRNAs and rescue constructs were seeded on Matrigel-coated Transwell filters. Invasion assays were performed as described in the Materials and Methods. (C) Sec5 expression analysis. 5'A prostate tumor cells transfected with indicated siRNAs and/or rescue constructs, were extracted and analyzed by SDS-PAGE and immunoblotting with antibodies to Sec5 (endogenous and ectopic proteins detected) or c-myc (ectopic Sec5 detected).