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. 2020 May 5;39:76. doi: 10.1186/s13046-020-01578-y

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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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — GLYR1 interacts with MLH1 and regulates its nuclear expression. a-b Correlation analysis of GLYR1 and MLH1 expression by immunohistochemical staining of paraffin-embedded human colorectal cancer tissue sections. Representative photographs of immunohistochemical staining are shown (200×, scale = 50 μm). The areas in the red and black panes indicate co-deficiency and co-expression, respectively. Pearson’s correlation analysis of GLYR1 and MLH1 according to immunohistochemical staining score, **r = 0.703, ***P < 0.001. c Co-immunoprecipitation was performed to validate the interaction between GLYR1 and MLH1 in SW480 cells. d Co-localization of GLYR1 (red) and MLH1 (green) in SW480 cells was assessed by laser-scanning confocal microscopy (× 2400, scale = 5 μm), DAPI (blue) stained nuclei. Error bars represent the mean ± SD of the Pearson’s correlation and overlap coefficient of GLYR1 and MLH1 (n = 5), *P < 0.05, **P < 0.01, ***P < 0.001. (E-F) Effect of GLYR1 knockdown on the expression and localization of MLH1 analyzed by immunofluorescence (× 2400, scale = 5 μm) and nucleoplasm separation assays. Grayscale values were normalized against GAPDH in the cytoplasm and histone H3 in the nucleus