Fig. 5

Effect of pertussis toxin (PTX) on 6-OAU treated microglial motility. a Microglia were pretreated with PTX (100 ng/mL) for 4 h and then exposed to 1 μM 6-OAU. Live imaging was performed for 30 min. The second and third rows show higher magnifications of cells indicated by arrows in the top row. Scale bar, 50 μm (top row) and 20 μm (other rows). b, c MetaMorph-based quantification of distance and velocity of cell migration. Cells were tracked over 30-min periods. Data represent mean ± SEM of 15 cells per group. d Rate of microglia undergoing membrane ruffling in each condition. Microglia were incubated with vehicle or 1 μM 6-OAU for 2 min in the presence or absence of PTX (100 ng/mL). Data represent mean ± SEM. In total, 150 cells from at least three independent experiments were used for quantification. e Microglia were cultured in 96-well plates and treated with 50 μM forskolin for 20 min after 6-OAU treatment. cAMP levels were determined using the cAMP EIA kit and presented as mean ± SEM of three independent experiments. *p < 0.05, ***p < 0.001