Figure 2.

Specificity of Aβ42-activated NFκB. Gel mobility-shift assay with nuclear extracts prepared from untreated astrocytes (lane 1), vehicle-control-treated astrocytes (lane 2), and 10 μM Aβ42-treated astrocytes (lanes 3–11). Cells were incubated for 12 hr. ³²P-labeled oligonucleotide shift probes used contained consensus NFκB response element (lanes 1–3 and 7–11), 50-fold molar excess of unlabeled AP2 (nonspecific competitor, NS) shift probe before ³²P-labeled NFκB shift probe (lane 4), approximately 50-fold molar excess of unlabeled NFκB (specific competitor, NFκB) shift probe before ³²P-labeled NFκB shift probe (lane 5), or ³²P-labeled mutant NFκB carrying a 1-bp substitution within the NFκB response element (lane 6). Polyclonal antibodies used to detect the indicated Rel family subunits are as indicated (lanes 7–11). NFκB activation complexes are indicated by A–D. Supershifted complexes are indicated by S and were detected only in samples incubated with either p65/RelA or p50 antibody.