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. 2017 Nov 28;7:16469. doi: 10.1038/s41598-017-16714-6

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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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Activation of Erk in the resealing epithelial cells in the ileum. (A) H&E staining images of the normal mouse ileum (left), and the damaged ileal epithelia at 24 h (middle) or 48 h (right) after ischemia. Bar, 50 μm. (B) Representative images of the damaged ileum 48 h after ischemia. A biosensor [green, denoted as Villin(+)] and the Keima protein [magenta, denoted as Villin(−)] and FRET/CFP and SHG images are depicted as described in Fig. 1B. (C) The schematic view of the damaged area and the three-dimensional projection image using FRET/CFP images. The colour bar indicates the FRET/CFP ratio in IMD with the lower and upper limits of the ratio range. (D) Comparison of the FRET/CFP ratio between cells in normal crypts (denoted as Normal) and resealing epithelial cells (denoted as Damaged) at z = 24 μm (p < 0.05; t-test). The graph shows the representative results from 9 independent experiments. The average FRET/CFP values for the 9 experiments were 0.714 (for cells in normal crypts) and 0.835 (resealing epithelial cells). (E) The cross sectional view of the three-dimensional reconstruction by 75 images taken at 1-μm intervals (Movie 2).