Figure 2. Lysophosphatidylcholine (LPC) mediates the enhancement of vesicle clustering by N-terminal acetylation of α-synuclein (α-syn).

(a) Scheme of single-vesicle clustering assay for the functional study of Ac-α-syn on vesicle clustering. Vesicles were prepared with different amounts of LPC, and were labeled with DiD or remained unlabeled, respectively. A saturated layer of unlabeled vesicles was immobilized on the imaging surface. Free DiD-vesicles were injected into the system with Ac-α-syn. Red laser illumination imaged the DiD-vesicles that clustered with unlabeled vesicles. The enhancement of LPC (b) and dioleoyl-phosphoserine (DOPS) (c) on single vesicle clustering count by Ac-α-syn and un-α-syn, respectively, was measured. Error bars are standard deviations from six random imaging locations in the same sample channel. *** indicates p-value <0.001, analysis by Student’s t-test.