Additional file 5: Figure S4. of Heparan sulfate proteoglycans mediate Aβ-induced oxidative stress and hypercontractility in cultured vascular smooth muscle cells

Aβ1-40 and Aβ1-42 remain mostly in soluble, monomeric form after 30 minute incubation in L-15 media. Conditioned media from Aβ1-40- (panels A, B) and Aβ1-42-treated VSMC (5 μM) was loaded onto a size exclusion column and fractionated via fast performance liquid chromatography (FPLC). Aβ-containing fractions were identified using enzyme-linked immunosorbent assays (ELISA) and higher-order Aβ species were separated from monomer (4 kDa) by SDS- (panels A, C) and native- (panels B, D) polyacrylamide gel electrophoresis (PAGE) with Western blotting. Aβ1-40 and Aβ1-42 peptides were detected using the anti-Human Aβ mouse 82E1 monoclonal antibody (1 μg/mL). To specifically differentiate monomers vs. oligomers in the media, we performed dot blot using oligomer-specific anti- Aβ antibody (A11; Panel E). (JPEG 75 kb)