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. 2020 Jun 9;26:56. doi: 10.1186/s10020-020-00182-2

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — iNOS is required for IRF-1 translocation to the nucleus and liver injury in I/R mice. a iNOS^+/+ and iNOS^−/− mice were used to generate I/R with 1 h ischemia and reperfusion as indicated. The nuclear and cytosolic proteins from the livers were analyzed by Western blot. b Livers from iNOS^+/+ and iNOS^−/− I/R mice (6 h reperfusion) were subjected to immunofluorescence staining. Representative images are shown in the comparison of IRF1 expressions between the iNOS^+/+ and iNOS^−/− mice. IRF1 is stained with FITC (green), and nucleus is stained with Hoechst dye (bis-benzimide) and is shown as blue color (upper). Moreover, to confirm the translocation of IRF1 to nucleus we used staining for IRF1 with Cy3 (red), and counterstaining for nucleus with SYTOX (green). Merging of the images shows the translocation to the nucleus of IRF1 as yellow color (lower). c ALT was detected in I/R iNOS^+/+ vs. iNOS^−/− mice at the indicated time points. I/R more likely induced ALT releases with a time-course dependent manner in iNOS^+/+ mice compared with that in iNOS^−/− mice, P < 0.0001. Data represent the mean ± SD, n = 5. d H&E staining of liver sections visualized in liver IRI in iNOS^+/+ vs. iNOS^−/−. Original magnification is × 100. e The necrotic areas were quantified with NIH ImageJ 2. Data are presented as mean ± standard deviation (n = 5, * P < 0.001 iNOS^+/+ vs. iNOS^−/− at each time point)