Figure 1

DJ-1 interacted with MAP1b light chain. A, GST-DJ-1 or GST was incubated with 6xhis-MAP1b-LC for 3 hrs and then was pulled down by GST beads, the beads were washed and SDS-PAGE followed by Coomassie blue staining was used to analyze the result. The result showed DJ-1 could bind to MAP1b-LC directly in vitro. B, Flag-MAP1b-LC and HA-DJ-1 were transfected into HEK293t cells for 36 hrs and the cell lysates were immunoprecipitated with Flag M2 beads or HA antibody conjugated beads. Both DJ-1 and MAP1b-LC were detected by western blot with HA or Flag antibody respectively. C and D, Co-localization of MAP1b-LC and DJ-1. Flag-MAP1b-LC and HA-DJ-1 were cotransfected into KEK293t (C) or SH-SY5Y cells (D) for 36 hrs before fixed and stained with rabbit anti-Flag and mouse anti-HA antibodies. E. The wild type or DJ-1 KO mouse brain lysates were immunoprecipitated with DJ-1 antibody, MAP1b-LC could be detected in the wild type mouse brain but not in that of the KO mouse. F, The primary cortical neuron was fixed and immuno-stained with DJ-1 and MAP1b-LC antibody.