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. 2019 Jul 19;10:3221. doi: 10.1038/s41467-019-11118-8

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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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Fig. 6 — Spatiotemporal FRAP analysis of Satb1 dynamics in the nucleus. a A schematic representation of the domain organization of Satb1 and the various domain constructs which were generated to study Satb1 dynamics. b Satb1 knockout lines were generated by CRISPR/Cas9 strategy. Immunofluorescence image of WT Vl3 3M2 cells and Vl3 3M2 cells where the native Satb1 has been knocked out (Satb1 green and DNA in red) c Examples of time versus fluorescence intensity carpets of high resolution spatiotemporal maps of fluorescence bleaching and recovery along scan lines bisecting a bleach spot in Satb1^−/− cells expressing exogenous eGFP fusions of FL Satb1 and different domain constructs. d The best-fit surface to a FL Satb1 spatiotemporal FRAP data. e, f The best fits of representative recovery profiles for N (e) and FL (f) are displayed at select times, with the color bar representing time in seconds. For estimates of spatiotemporal FRAP fit parameters, as well as total number of cells analyzed per condition, see tables 1 and 2. Scale bar is 5 µm. See source data for analysis and Supplementary Code 1 for custom analysis codes