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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. 4 — Downregulation of GLYR1 inhibits cell differentiation. a Representative photographs of GLYR1 immunohistochemical staining (× 200, scale = 50 μm) in cases with different degrees of differentiation. Spearman’s correlation analysis of GLYR1 expression (Low, Medium, High) and tumor grade (G1, G2, G3),** R = 0.319, ***P < 0.001. b Relative mRNA expression of cell differentiation-related genes (p21, p27, E-cadherin, CK20, CD133, NANOG) and GLYR1 was measured in SW480 and SW620 cells after ATRA treatment by qRT-PCR; GAPDH was used as an internal control. c Western blot analysis of the expression of E-cadherin, CD133, GLYR1, and p21 in SW480 and SW620 cells after ATRA treatment. d Relative mRNA expression of cell differentiation-related genes (p21, p27, E-cadherin, CK20, CD133, NANOG) was measured in GLYR1-knockdown SW480 and SW620 cells by qRT-PCR; GAPDH was used as an internal control. e Western blot analysis of the expression of E-cadherin, CD133, and p21 in GLYR1-knockdown SW480 and SW620 cells. f Immunofluorescent staining of the expression of p21, E-cadherin, and CD133 in GLYR1-knockdown SW480 cells (× 2400, scale = 5 μm); DAPI (blue) stained nuclei. Error bars represent the mean ± SD (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001