Sr a1

Rat-derived primary microglia were pre-incubated with scavenger receptor-targeting treatments, including either 10 µg/mL SRA1 (R&D) and CD36 (Abcam) antibodies or NPs for 24 hours, followed by 24 hour co-incubation with 5 µM monomeric ASYN (rPeptide), ^{48 (link)} fluorescently labeled with DyLight 594 nm (Dy594) microscale antibody labeling kit (Thermo Fischer) or 5 µM monomer-equivalent fibrillar ASYN. For fibrillar ASYN studies, cells were fixed prior to characterization by immunocytochemistry. For monomeric ASYN studies using live cells, extracellular non-internalized synuclein fluorescence quenched by adding 0.5 mg/mL trypan blue (VWR) for 30 minutes, followed by two washes with PBS ^{49 (link)}. Live cells were imaged on a Leica SP2 confocal microscope using a 40 x dry objective. Intracellular fluorescence quantification was performed using NIH-ImageJ software (http://rsb.info.nih.gov/ij/), by measuring mean grey value in cells segmented by applying the same fluorescence thresholds to all collected images. For untreated cell controls, fluorescence images were segmented based on cell boundaries visualized in brightfield images.