Figure 5.

Analysis of cell migration and invasion of glioma cell lines upon manipulating the gene expression of integrin α3. (A) Extracts of U87, SNB19, or U251 cells transiently transfected with integrin α3 (ITGA3) or empty plasmid vector (Mock) were immunoblotted with antibodies against ITGA3 or β-actin. (B) Transfected cells were plated on membranes coated with fibronectin (upper panel) or laminin (lower panel) in a chemotaxis chamber. Bar: mean±s.e. (n=6). Asterisk indicates a significant difference from the control at a P-value of <0.05. (C) U87 cells transfected with integrin α3 (ITGA3) or empty plasmid vector (Mock) (upper panel) and treated by siRNA for integrin α3 (siITGA3-1 and siITGA3-2) or luciferase (lower panel) were plated on membranes coated with Matrigel in a chemotaxis chamber. Cells were allowed to migrate to the underside surface of membranes for 24 h; invading cells were stained with Diff-Quick and the absorbance was measured at 490 nm, as described in Materials and Methods. The mean absorbance (490 nm) value from mock or control cells in each figure is shown as 1. Bar: mean±s.e. (n=6). Single and double asterisks indicate significant differences between mock and control at P-values of <0.05 and <0.01, respectively.