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. 2020 Jun 9;219(7):e201911047. doi: 10.1083/jcb.201911047

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© 2020 Shi et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 1. — The FIP200NTD scaffolds the ULK complex as a homodimer. (A) Domain diagram of the ULK complex proteins. (B) Pulldown assay of ULK1, ATG13, and ATG101 with FIP200NTD (N-640) and FIP200CTD (636-C). Strep-Tactin resin was loaded with MBP-ULK1:Strep-ATG13:Strep-ATG101 complex to pull down FIP200(N-640)–MBP and FIP200(636-C)–MBP. The pulldown results were visualized by SDS-PAGE and Coomassie blue staining. (C) NSEM 2D class averages for FIP200NTD (top), N-terminal MBP–tagged FIP200NTD (middle), and C-terminal MBP-tagged FIP200NTD (bottom). Densities corresponding to MBP tags are labeled with yellow arrows. Scale bar is 20 nm. (D) MALS and SEC trace of FIP200NTD shows the predicted and measured molecular weight of the dimeric FIP200NTD. MW, molecular weight.