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. 2018 Nov 8;9:4684. doi: 10.1038/s41467-018-07037-9

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Characterization of recombinant Chlamydomonas IFT172. a Predicted domain architecture of IFT172 from Chlamydomonas. The predicted WD40 repeats (in yellow) and TPR (tetratricopeptide repeat) domain (in blue) were defined by HHpred. IFT172 shows a similar domain organization to COPI/II and clathrin. COPI β’ from Saccharomyces cerevisiae is shown for comparison. b Size-exclusion chromatography (SEC) profile of recombinant full-length IFT172 and Coomassie-stained SDS-PAGE analysis of recombinant IFT172 after size-exclusion chromatography. c Dynamic light scattering (DLS) measurement of IFT172 shows a uniform particle size distribution of IFT172 with an average diameter of ~120 nm with 100% mass. d Negative stain EM micrograph of IFT172 showing globular oligomer formation. Scale bar: 100 nm. e SEC profile of IFT172 proteolyzed products after limited-proteolysis (left) and SDS-PAGE of the peak fractions from SEC (right). f Negative stain EM micrograph of the fraction from the void peak in e shows round particles with smooth surface. Scale bar: 100 nm. g Negative stain EM micrograph of the liposomes from Folch fraction I as a control. Scale bar: 100 nm. h Scheme describing the experimental procedure of e and b