Figure 5

Effects of chemical kinase inhibitors on AAH protein expression and directional motility in IGF-I stimulated SH-Sy5y cells. Subconfluent cultures were serum starved over night, then stimulated with IGF-1 and treated with vehicle (Con), or a chemical inhibitor of Akt, PKA (H-89), GSK-3β (LiCl), Erk MAPK (PD98059-PD9), Cdk-5 (Roscovitine-Rosc), or p38 MAPK (SB202190-SB2). Inhibitor concentrations are listed in Table 1. After 24 hours of growth factor stimulation, cells were harvested to measure AAH protein by (A) Western blot analysis with (B) digital imaging, or (C) they were analyzed directly in 96-well micro-cultures using the MICE assay. (D) Directional motility was measured using the ALMI assay (see Methods). (A-lower panel) For Western blot controls, the blots were stripped and re-probed with monoclonal antibodies to β-actin. The graphs depict the mean ± S.D. of results. Statistical comparisons were made using ANOVA and post hoc Fisher LSD tests (*P < 0.001 relative to control).