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. 2019 Oct 23;9:15244. doi: 10.1038/s41598-019-51639-2

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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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Colon homeostasis is impaired in epithelial EP4-deficient mouse. (A) Schema of recombination in Villin-Cre; EP4^flox/flox mice. (B) Representative microscopic view of isolated crypts. Scale bars = 100 μm. (C) qRT-PCR analysis of EP4 mRNA levels in Villin-Cre (n = 5) and EP4 cKO (n = 5) colon crypts from the mice at 8 weeks of age. (D) Hematoxylin and eosin (H,E) staining of colon for Villin-Cre and EP4 cKO mice. Scale bars = 50 μm. (E) Crypt length in Villin-Cre and EP4 cKO mouse colons (n = 4). (F) Cell size of colonic epithelial cells located in the top of colon crypts in Villin-Cre and EP4 cKO mice (n = 3). (G,H) Alcian Blue staining and quantification in the colons from Villin-Cre and EP4 cKO mice at 8 weeks of age (n = 3). Scale bars = 50 μm. (I) Muc2 staining of colons in Villin-Cre and EP4 cKO mice at 8 weeks of age. Scale bars = 50 μm. (J) qRT-PCR analysis of Muc2 mRNA levels in Villin-Cre and EP4 cKO mice (n = 5). (K) Chromogranin A staining of colon in Villin-Cre and EP4 cKO mice at 8 weeks of age. Scale bars = 50 μm. (L) Quantification of (K) (n = 3) and mRNA expression levels of Chromogranin A in Villin-Cre and EP4 cKO mice (n = 4) analyzed by qRT-PCR on colon crypts. (M) Dclk1 staining of colon in Villin-Cre and EP4 cKO mice at 8 weeks of age. Scale bars = 50 μm. (N) Quantification of (M) (n = 3) and mRNA expression levels of Dclk1 in Villin-Cre and EP4 cKO colon crypts (n = 4) analyzed by qRT-PCR. Results are shown as mean ± SEM. ***P < 0.005.