Figure 2

PKA may mediate the effects of cilostazol and cAMP. (a) Cilostazol and cAMP activate PKA in astrocytes. Western blot analysis of p-PKA in cells treated with 10 μM cilostazol (+Cilo) or 300 μM cAMP alone (+cAMP), or together with 10 μM H-89 (+PKAi), a PKA inhibitor, for 1 hour. H-89 reversed cilostazol- or cAMP-induced increases in p-PKA levels. Bars denote the ratio of p-PKA bands to corresponding total PKA bands. Data are presented as means ± SEM (n = 7; * denote P < 0.05, ** denote P < 0.01 compared with CTL, Cilo, cAMP or PKAi, Two-tailed Student’s t-test for 2 comparisons). (b) DND-189 fluorescence in astrocytes before (CTL) and after a 60-minute exposure to BafA1 (BA), BafA1 plus cilostazol (BA + Cilo), BafA1 plus cAMP (BA + cAMP) or BafA1 plus forskolin (BA + forsk), in the absence or presence of PKAi (BA + PKAi) (Scale bar, 20 μm). (c) Bars indicate relative changes in the fluorescence intensity. Values for individual bars were normalized to control values (mean ± SEM; n = 4; * denote P < 0.05 compared with BA, Cilo, cAMP, forsk or PKAi; Two-tailed Student’s t-test for 2 comparisons).