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. 2017 Dec 1;7:16741. doi: 10.1038/s41598-017-16842-z

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© The Author(s) 2017

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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Expression of QUAD opsin. (A) Protein expression. WT and QUAD opsins were expressed as thermostable, FLAG-tagged variants in HEK293S GnTI⁻ cells, extracted with dodecyl-β-D-maltoside (DDM) and purified by FLAG affinity chromatography. An SDS-PAGE Coomassie-stained gel is shown of samples obtained from the same number of cell equivalents. (B) Fluorescence size exclusion chromatography (FSEC) analysis. GFP-tagged WT and QUAD opsins were expressed in HEK293S GnTI⁻ cells, and a DDM extract of the cells was analyzed (without purification of the expressed proteins) by FSEC. Both proteins display mono-disperse profiles (the trace for QUAD opsin is vertically displaced from that of WT opsin for clarity). Elution positions of dextran (void volume marker) and albumin are shown.