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. 2017 Aug 16;7:8450. doi: 10.1038/s41598-017-08493-x

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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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Labelling of TubA1B gene through CRISPR-Cas9 genome editing in 3 cell lines. (a) mEGFP tagging of TubA1B was compared to antibody based labelling in Hel 92.1.7, A549 and Hek293T cells. In all cases, gene editing results in the expression of an mEGFP α-tubulin fusion protein under endogenous control mechanisms (10 μm scale). (b) mEos 3.2 tagged TubA1B allows for the endogenous expression of a photoswitchable tag, and thus PALM imaging. Resolution is similar to that observed for dSTORM images in Hel 92.1.7 cells, and is dependent on the expression level of the mEos 3.2 fusion (c) Detection metrics from ThunderSTORM reconstructions evidence no significant difference between mEos 3.2 imaging in Hel 92.1.7 cells and dSTORM, while the lower expression level in A549 and Hek293T cells results in significantly reduced localization precisions (xy uncertainties) and number of localizations. Bars represent mean ± SD or Mode ± SD. (n = 3). Data were analysed using a 2-way ANOVA with multiple comparisons (Tukey’s). (10 μm scale in whole cell images, 1μm scale in cropped fields). Full sized images are supplied in Supplementary Fig. 4.