Fig. 1.

Analysis of exosomes released by SH-SY5Y. Characterisation of exosomes isolated from conditioned medium (CM) from normal and alpha-synuclein over-expressing SH-SY5Y cells: a, Electron micrograph of phosphotungstic acid-stained exosomes; scale bar = 200 nm. b, NTA analysis of particle diameters in exosome preparations from CM from normal (green line) or alpha synuclein expressing (red line) SH-SY5Y cells. c, Western blot analysis of exosome markers (LAMP-1, Alix and flotillin-1) and alpha-synuclein (alpha-synuclein and HA). d, Conditioned medium from alpha synuclein expressing cells was immunoprecipitated (IP) with anti-tubulin or anti-flotillin-1 antibodies. All the eluted IP protein and 40% (15 μg) of the flow through (FT) were loaded and probed with anti-LAMP-1 and anti-alpha-synuclein antibodies. e, ELISA quantitation of alpha-synuclein levels in the flow through after IP with flotillin-1 presented as a percentage of the level in the CM.