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. 2020 Oct 19;11:5273. doi: 10.1038/s41467-020-18970-z

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© The Author(s) 2020

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 — a Dehydrodolichyl diphosphate synthesis reaction scheme. The IPP moiety is colored red. The hcis-PT subunits DHDDS and NgBR are schematically drawn and colored blue and yellow, respectively. b Representative SEC-MALS analysis of the purified shcis-PT. The black and red curves indicate UV absorption and molecular mass, respectively. The experiment was performed twice using two different protein batches. Inset: SDS-PAGE analysis of the purified complex. Left lane: molecular weight marker, right lane: purified shcis-PT. Molecular weights (kDa) are indicated. Source data are provided as a Source Data file. c Native ESI-MS spectrum obtained using low activation conditions. The main distribution corresponds to the heterotetramer (charge states 21–26). Inset: SDS-PAGE analysis of the complex following glutaraldehyde cross-linking. Left lane: molecular weight marker, middle lane: purified shcis-PT without glutaraldehyde, and right lane: purified shcis-PT with glutaraldehyde. Molecular weights (kDa) are indicated. The experiment was performed twice using two different protein batches. The oligomeric state represented by each state is illustrated using the schematic presentation from panel (a). Source data are provided as a Source Data file. d In vitro activity of purified shcis-PT assessed as IPP incorporation. Experiments were performed as described in the “Methods” section. Data are presented as mean ± SEM (n = 3 independent experiments). Source data are provided as a Source Data file.