Figure 3.
Transient manipulation of tyrosine kinase B (TrkB) in head and neck squamous cell carcinoma (HNSCC) alters ligand-mediated migration and invasion. (a) OSC19 cells were transiently transfected with siRNA constructs, and lysates from subconfluent cells were analyzed by SDS–PAGE with the indicated antibodies. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) knockdown was assayed as a positive control. (b) OSC19 cells were transiently transfected with small interfering RNA (siRNA) against TrkB (construct 3) or a non-targeted siRNA (inset), and seeded on Transwell migration plates for 24 h in the presence or absence of BDNF (100 ng/ml). Cells were counted in five high-powered fields and analyzed for differential migration with ImagePro. Experiments were performed in triplicate and repeated three times. Columns, number of migrated cells in the presence or absence of BDNF; bars, s.e.m.; NS, not significant; magnification: × 100. (c) OSC19-Luc cells were transiently transfected with siRNA against TrkB (construct 3) or a non-targeted siRNA, and seeded on Matrigel invasion plates for 24 h. Cells were counted with an inverted fluorescence microscope in five high-powered fields and analyzed for differential invasion with ImagePro. Experiments were performed in triplicate and repeated three times. Columns, relative invasion of cells; bars, s.e.m, magnification: × 100.
