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. 2018 May 8;9:973. doi: 10.3389/fimmu.2018.00973

Figure 4.

Figure 4

Immunohistochemistry of colons. (A) In situ hybridization (RNAScope®) of Lgr5 in murine colon. Nuclei were counterstained with hematoxylin, scale bars indicate 100 μm (B) Lgr5 mRNA expression relative to housekeeping gene EF in colon tissue from wild-type (WT) and NoxO1 knockout (NoxO1−/−) mice. n = 7. (C) Immunohistochemistry staining of mitosis marker phospho-Histone 3 (pH3). Representative pictures and analysis of pH3-positive cells per crypt. n = 3, *p < 0.05. Scale bars indicate 200 μm. (D) Multiplex immunohistochemistry of murine colon. Tissue was stained for DAPI (white), alpha-smooth muscle actin (aSMA, green), Ki67 (blue), cleaved Caspase 3 (cCas3, magenta), and pan-cytokeratin (Pan-CK, orange); scale bars indicate 100 μm. Positive cell fractions for indicated phenotypes in the mucosa were analyzed using the in Form 2.0 software. n = 5, *p < 0.05.