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. 2019 Dec 30;9:20369. doi: 10.1038/s41598-019-56814-z

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© The Author(s) 2019

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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Relative single channel glycerol and urea permeabilities of AQP4 selectivity filter mutants. (A) Cell swelling rate constants of HEK293 cells transfected with AQP4 mutants, calculated from calcein fluorescence time-series data. (B) Cell-surface expression of AQP4 mutants in HEK293 cells measured by cell-surface biotinylation and neutravidin ELISA. (C,D) Representative calcein fluorescence timeseries from HEK293 cells transfected with AQP4 H201A or H201G and exposed to iso-osmotic solutions containing 150 mM of urea or glycerol. (E) Cell swelling rate constants from (A) above normalized to surface expression and then normalized to AQP3 glycerol permeability. This gives a relative single channel permeability in units in which AQP3 glycerol permeability is 1. (F) Normalized permeabilities for the analogous mutants of AQP1 for comparison. Panels A, B, D and F are adapted from P. Kitchen PhD thesis³⁵.