Figure 5

Knockdown of endogenous SNAI2 /Slug decreases glioblastoma migration and invasion. A) Human U87 glioblastoma cells were transduced to over express an shRNA directed against SNAI2/Slug (shSlug) or a control vector. Total RNA was then collected and analyzed for SNAI2/Slug mRNA expression by Taqman Real-time PCR. shSlug decreased SNAI2/Slug mRNA expression by approximately 60%. shRNA-mediated knockdown of Slug protein was confirmed by Western blot (right panel). B) Transwell migration assay for human U87 glioblastoma cells overexpressing an shRNA directed against SNAI2/Slug or a control vector. 10% serum was used as a chemoattractant. The cells that migrated through the membrane were stained with H&E and counted under direct microscopy. shSlug-expressing glioblastoma cells migrated slower than the control cells (asterisk indicates P < 0.0002, t-test). C) 3-dimensional Matrigel assay for human U87 glioblastoma cells overexpressing an shRNA directed against Slug or a control vector. 24 hours after plating, cells that invaded through the matrix were stained and counted under direct microscopy. Data shown are mean ± SEM. Glioblastoma cells overexpressing shSlug showed less invasion than control cells (asterisk indicates P < 0.0001, t-test).