Fig. 6

Dasatinib reduces LPS-induced cytosolic and nuclear p-STAT3 levels in BV2 microglial cells. a BV2 microglial cells were treated with vehicle (1% DMSO) or dasatinib (250 nM) for 30 min followed by PBS or LPS (1 μg/ml) for 5 h 30 min, and western blotting of the cytosolic fraction was performed with anti-p-STAT3 (Ser727) and anti-β-actin (as a cytosolic marker) antibodies. b Quantification of the data in a (con, n = 12, LPS, n = 12, dasatinib+LPS, n = 12). c BV2 microglial cells were treated with vehicle (1% DMSO) or dasatinib (250 nM) for 30 min followed by PBS or LPS (1 μg/ml) for 5.5 h, and western blotting of the nuclear fraction was conducted with anti-p-STAT3 (Ser727) and anti-PCNA (as a nuclear marker) antibodies. d Quantification of the data in c (con, n = 21; LPS, n = 21; dasatinib+LPS, n = 21). e BV2 microglial cells were treated with vehicle (1% DMSO) or dasatinib (250 nM) for 30 min followed by PBS or LPS (1 μg/ml) for 5.5 h, and immunostaining was performed with anti-p-STAT3 (Ser 727) and anti-CD11b antibodies. f Quantification of the data in e (con, n = 437; LPS, n = 527; dasatinib+LPS, n = 338). g BV2 microglial cells were treated with S3I-201 (STAT3 inhibitor, 50 μM) or vehicle (1% DMSO) for 30 min, treated with vehicle (1% DMSO) or dasatinib (250 nM) for 30 min, and treated with PBS or LPS (1 μg/ml) for 5 h. RT-PCR was then performed to measure proinflammatory cytokine levels. h, i Quantification of the data in g (COX-2: con, n = 26; LPS, n = 26; dasatinib+LPS, n = 26; S3I-201+LPS, n = 26; S3I-201+dasatinib+LPS, n = 26; and IL-6: con, n = 30; LPS, n = 30; dasatinib+LPS, n = 30; S3I-201+LPS, n = 30; S3I-201+dasatinib+LPS, n = 30). One-way ANOVA with Tukey’s post hoc test was used to analyze significant differences. *p < 0.05, **p < 0.01, ***p < 0.001