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. 2019 Aug 6;12:189. doi: 10.3389/fnmol.2019.00189

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Copyright © 2019 Du, Wu, Luo, Lu and Ma.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Preparation of recombinant α-syn-98 aggregates. (A) Quality of purified recombinant fusion protein α-syn-98-GST was estimated by SDS-PAGE, and gels were stained with Coomassie brilliant blue. Lane 1: total protein obtained from E. coli BL21 expressing recombinant plasmid (pGEX-5X-1-α-syn-98). Lane 2: α-syn-98-GST protein purified from lysates using GST fusion protein purification beads (BeaverBeads^TM GSH). (B) Western blot analysis of purified α-syn-98. Mouse monoclonal antibody specific for α-syn (ALX-804-656, ENZO) was used. (C) The aggregates formation monitored by ThT assay for up to 8 days. (D) The proteinase K resistance (5 μg/ml) of recombinant α-syn-98 aggregates analysis by SDS-PAGE, and gels were stained with Coomassie brilliant blue. (E) EM determined the conformation of α-syn-98 aggregates obtained by incubation for 6 days at 37°C.