In vitro dephosphorylation

100 μg of α-syn GST fusion protein on beads were resuspended in alkaline

phosphatase (ALP) buffer supplemented with 250 units of ALP (Thermo Scientific

Cat# EF0651).

The mixes were incubated for 60 min or 180 min at 37 °C with

orbital shaking (Eppendorf ThermoMixer F1.5).

Samples were then analyzed by

NuPAGE and immunoblotted or alternatively used in pull-down experiments with

brain lysates.