Fig. 6

Confirmation of ANT1 interaction with α-synuclein in vitro. M, Page Ruler prestained protein ladder (Fermentas); 1, cell lysate of E.coli containing over-expressed α-synuclein; 2, eluent fraction-1 from Ni-NTA affinity chromatography (purified α-synuclein); 3, eluent fraction-2 from Ni-NTA affinity chromatography (purified α-synuclein); 4, eluent fraction from pull-down assay (interaction proteins); 5, cell lysates of mouse brains. a SDS-PAGE analysis of α-synuclein expressed in E.coli. The gel was stained with Coomassie blue. b Western blot analysis of the purified α-synuclein using monoclonal anti-6XHis antibody as the probe. The soluble α-synuclein was purified by Ni-NTA affinity chromatograph. c SDS-PAGE analysis of interaction proteins with α-synuclein. The interaction proteins were pulled down using Ni-NTA magnetic agarose beads as the medium and the purified α-synuclein as the bite. The gel was visualized by silver staining. d Western blot analysis of interaction proteins with α- synuclein against anti-ANT1 monoclonal antibody. e Profiles of the identified ANT1 in Mus musculus by LC-MS/MS