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Fig. 1 | Molecular Neurodegeneration

Fig. 1

From: Initiation and propagation of α-synuclein aggregation in the nervous system

Fig. 1

Human α-synuclein fibrils were labeled with Alexa-488. To visualize fibrils at the cell surface, primary hippocampal neurons were incubated at 4 °C for 30 min with Alexa-488 labeled fibrils, rinsed, and images were captured. To visualize internalized Alexa-488 labeled fibrils and oligomers, primary hippocampal neurons were pre-incubated with the fibrils or oligomers for 30 min at 4 °C and then incubated for 15 min at 37 °C. The neurons were rinsed and incubated with trypan blue to quench extracellular Alexa-488 label and to visualize intracellular fibrils and oligomers. When trypan blue binds to proteins on the cell surface, it fluoresces at 560 nm, which is shown in the images as magenta. The right panel shows quantitation of internalization of unsonicated, long fibrils (F-L), sonicated 50 nm fibrils (F-s) and oligomers (O). From Froula JM, Castellana-Cruz M, Anabtawi NM, Camino JD, Chen SW, Thrasher DR, Freire J, Yazdi AA, Fleming S, Dobson CM, et al.: Defining alpha-synuclein species responsible for Parkinson’s disease phenotypes in mice. The Journal of biological chemistry 2019, 294:10392–10,406

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