Figure 2. Oligomeric Aβ1-42 stimulated increased microglial protein phospho-tyrosine levels that were attenuated by the tyrosine kinase inhibitor, dasatinib.
(A) Primary microglia were vehicle treated (v), or stimulated for 5 min with 0.1μM, 1μM and 5μM Aβo. Cells lysates were resolved by 10% SDS-PAGE and Western blotted using anti-phospho-Tyr (4G10), anti-pSrc, anti-pLyn antibodies or anti-α-tubulin, anti-Src and anti-Lyn (loading controls) antibodies. A Representative blot from 5 independent experiments is shown. (B) Densitometric analyses of the Western blots was performed normalizing protein phospho-tyrosine, pSrc and pLyn levels against their respective α-tubulin, Src and Lyn controls and averaging +/−SD. Percent fold changes in phospho-tyrosine levels were plotted (*p< 0.05 from vehicle). (C) Primary microglia were stimulated for 5 min with 1μM Aβo with or without 30 min pretreatment with 100pM dasatinib (d). A representative Western blot from 3 independent experiments is shown. (D) Densitometric analyses of Western blots were performed for protein phosphotyrosine levels normalized to their respective α-tubulin control (*p<0.05 from vehicle and dasatinib, **p<0.01 from Aβ+dasatinib).