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. 2021 Apr 22;12:2363. doi: 10.1038/s41467-021-22329-3

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© The Author(s) 2021

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. 4 — Enrichment of heterologous membrane proteins in S12 and S30 extracts was quantitated using α-FLAG western blots against the heterologous proteins for: a glycosylation enzymes, b signal transduction proteins, and c a cytosolic sfGFP control with no transmembrane helices. On each western blot, left lanes are S12 extracts and right lanes are S30 extracts. Black arrows indicate the membrane protein of interest. Molecular weight (kDa) from protein ladder standards are indicated to the left of each blot. Protein names and enrichment ratios of bands (S12/S30) are shown directly below each blot. All blots are representative of n = 3 biologically independent extracts. Cartoons depict the transmembrane topology for each protein. See Supplementary Table 1 for taxonomical origin, transmembrane topology, functions(s), theoretical size, and UniProt ID. Full western blot images for panels (a–c) are available in Supplementary Fig. 6 and Source Data file. d Fluorescence chromatograms of SEC analysis of extracts probed with a fluorescent α-FLAG antibody. Strains used to prepare extracts were enriched with no membrane protein (gray trace), PglB (dark purple trace), or PglO (light purple trace). Characteristic vesicle elution fraction from 3 independent experiments is highlighted in gray. Source data for all panels are provided as a Source Data file.