Dasatinib dose-dependently attenuated active, phospho-Src kinase levels in the BV2 cell line and primary microglia. (A) Microglial BV2 cells were vehicle treated (v), or treated with 1 nM, 10 nM, 100 nM and 1 μM dasatinib for 30 minutes. Cells lysates were resolved by SDS-PAGE and Western blotted using anti-phospho-Src, and Src antibodies. (B) Densitometric analyses of the Western blots were performed normalizing active-Src levels against their respective Src controls and averaging +/−SEM. Blots are representative of six independent experiments. Primary microglia were DMSO vehicle treated (veh), or stimulated for 5 minutes with 10 μM Aβf in the presence or absence of 30-minute pretreatment of 100 nM dasatinib. (C) Cells lysates were Western blotted using anti-pSrc or Src (loading control) antibodies and anti-pLyn or Lyn (loading control) antibodies. A representative blot from six independent experiments is shown. Densitometric analyses of the Western blots was performed normalizing (D) active, phospho-Src levels against Src controls and (E) active, phospho-Lyn levels against Lyn controls and averaging +/−SEM. Percent fold changes in phospho-Src and phospho-Lyn levels were plotted. (*P <0.05 vs. vehicle, dasatinib, **P <0.01 vs. Aβ + dasatinib for pSrc and *P <0.01 vs. Aβ for pLyn).