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. 2018 May 2;8:6911. doi: 10.1038/s41598-018-25083-7

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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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The internal optical signal size decreases during repeated patch clamp measurements in HEK293 cells. A HEK 293 cell (same cell that was illustrated in Figs 1 and 2) expressing Aahn was voltage-clamped. The internal signal, but not the external signal, decreases with repeated trials. (A) The resting light image of the Aahn expression. (B,C) Frame subtraction images for the first 4 trials and the last 4 trials (trials 21–24) in response to the 100 mV depolarization pulse. The internal signal area is somewhat smaller in the last group of 4 trials. (D) The optical traces of internal and external signals for the six sets of four trials are shown. Each trace is the average of 4 trials without temporal filtering. Images were recorded at a frame rate of 500 fps. The size bar represents 10 μm.