Membrane binding, internalization, and sorting of alpha-synuclein in the cell

Abstract

[...]samples were sonicated (10 pulse/second) and immunoblotting analysis were performed as described above. [...]cells were stained with Hoechst 33,258 (Life Technologies- Invitrogen, Carlsbad, CA,USA) (1:5000 in DPBS) for 5 min, washed again and then fixed with Mowiol (Sigma Aldrich, St. Louis, MO, USA) for epifluorescence and confocal microscopy. [...]to confirm the functional involvement of Rab5A on the internalization of aSyn, we used a mutant in which the GTPase activity is deregulated, resulting in permanent activation - constitutively active mutant - Rab5ACA-GFP (Fig. 6g-i). [...]impaired autophagic degradation of aSyn could lead to the accumulation of aggregation-competent species [19].