Figure 2

Aβ1–42 stimulated a conformation-specific MAP and tyrosine kinase signaling response along with increased COX-2 and CD68 protein levels. Primary mouse microglia were unstimulated (control) or stimulated with 20 μM Aβo or 20 μM Aβf. (A) Cells were lysed after 5 min with RIPA buffer and lysates were separated by 10% SDS-PAGE and Western blotted with anti-phosphotyrosine antibody (4G10), anti-phospho-JNK antibody, anti-JNK antibody (loading control), anti-phospho-ERK antibody, anti-ERK2 antibody (loading control), anti-phospho-p38 antibody, anti-p38 antibody (loading control), anti-phospho-Lyn antibody, or anti-Lyn antibody (loading control). (B) Cells were also lysed after 5 min with RIPA buffer and Syk was immunoprecipitated. Immunoprecipitates were separated by 7% SDS-PAGE and Western blotted with anti-phosphotyrosine antibody (4G10) and anti-Syk antibody. Arrowheads differentiate specific immunoreactivity from IgG heavy chain. (C) Cells were lysed after 24 hrs with RIPA buffer. Cell lysates were separated by 10% SDS-PAGE and Western blotted with anti-COX-2 antibody, anti-CD68 antibody, anti-CD45 antibody and anti-ERK2 antibody (loading control). Antibody binding was visualized by chemiluminescence. Blots are representative of at least three independent experiments.