Fig. 4

Dasatinib significantly suppresses LPS-induced AKT and/or ERK phosphorylation. a BV2 microglial cells were treated with vehicle (1% DMSO) or dasatinib (250 nM) for 45 min followed by PBS or LPS (1 ug/ml) for 45 min, and western blotting was performed with anti-p-ERK and anti-ERK antibodies. b, c Quantification of the data in a (p-ERK and ERK: con, n = 18; LPS, n = 18; dasatinib+LPS, n = 18). d BV2 microglial cells were treated with vehicle (1% DMSO) or dasatinib (250 nM) for 45 min followed by PBS or LPS (1 ug/ml) for 45 min, and western blotting was conducted with anti-p-AKT and anti-AKT antibodies. e, f Quantification of the data in d (p-AKT: con, n = 8; LPS, n = 8; dasatinib+LPS, n = 8 and AKT: con, n = 17; LPS, n = 17; dasatinib+LPS, n = 17). g BV2 microglial cells were treated with dasatinib (250 nM) or vehicle (1% DMSO) for 45 min followed by LPS (1 ug/ml) or PBS for 45 min, and immunocytochemistry was conducted with anti-p-ERK and anti-CD11b antibodies. h Quantification of the data in g (p-ERK: con, n = 239; LPS, n = 243; dasatinib+LPS, n = 212). i BV2 microglial cells were treated with dasatinib (250 nM) or vehicle (1% DMSO) for 45 min followed by LPS (1 ug/ml) or PBS for 45 min, and immunocytochemistry was conducted with anti-p-AKT and anti-CD11b antibodies. j Quantification of the data in i (p-AKT: con, n = 599; LPS, n = 654; dasatinib+LPS, n = 601). k Primary microglial cells were treated with vehicle (1% DMSO) or dasatinib (250 nM) for 45 min followed by PBS or LPS (1 ug/ml) for 45 min, and western blotting was performed with anti-p-ERK and anti-ERK antibodies. l, m Quantification of the data in k (p-ERK and ERK: con, n = 3; LPS, n = 3; dasatinib+LPS, n = 3). n Primary microglial cells were treated with vehicle (1% DMSO) or dasatinib (250 nM) for 45 min followed by PBS or LPS (1 ug/ml) for 45 min, and western blotting was conducted with anti-p-AKT and anti-AKT antibodies. o, p Quantification of the data in n (p-AKT and AKT: con, n =3; LPS, n = 3; dasatinib+LPS, n = 3). One-way ANOVA with Tukey’s post hoc test was used to analyze significant differences. *p < 0.05, **p < 0.01, ***p < 0.001