Nature research paper: Medin co-aggregates with vascular amyloid-β in Alzheimer’s disease
. The numbers of plaque-associated PU.1-positive microglia and GFAP-positive astrocytes were normalized to Congo Red-positive area for every individual plaque. Microglial nuclei were exclusively counted in the two-fold radius of plaque diameter. GFAP-positive cell area was calculated by measuring the radius of GFAP-positive cell staining per plaque.
ThT fluorescence aggregation assay, and electron microscopy of recombinant medin or amyloid-β co-aggregation and human aorta extracts ThT fluorescence assays were carried out on a Flexstation 3 microplate reader . Experiments were carried out in sealed 96-well, black-walled, clear-bottomed microplates . Data were recorded every 5 min using bottom read mode, with excitation at 440 nm and emission at 490 nm.
ThT fluorescence assays were carried out on an Fluostar Omega microplate reader . Experiments were carried out in sealed 96-well, half-area flat clear-bottomed microplates . Data were recorded every 10 min using bottom read mode, with excitation at 440 nm and emission at 490 nm. Prior to ThT assay experiments, lyophilised samples of Met-Aβ were suspended for 1 h at ambient conditions in 7 M guanidinium chloride in 50 mM Tris , then centrifuged for 5 min.
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