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. 2016 Sep 21;7:12887. doi: 10.1038/ncomms12887

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Copyright © 2016, The Author(s)

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(a–c) Conditional knockout (cKO) of parafibromin results in decreased expression of Wnt and Notch targets in the intestine. CD44 immunohistochemistry (a), Sox9 immunofluorescent staining (b) and Hes1 immunohistochemistry (c) of control and parafibromin cKO colons. The right panels show higher-magnification images. Scale bars, 20 μm. (d,e) Parafibromin is essential for the intestinal epithelial cell proliferation. Ki-67 immunohistochemistry (d) and number of Ki-67-positive cells per crypt (e) (n=4 animals per group. Means±s.d.; ^**P<0.01; a two-tailed unpaired Student's t-test). Scale bar, 20 μm. (f,g) Effect of parafibromin cKO on enterocyte differentiation. Haematoxylin and eosin (H&E) staining (f), and Villin immunohistochemistry (g) of colonic epithelial tissues from control and parafibromin cKO mice. (h) Parafibromin is essential for the maintenance of intestinal epithelium. H&E staining was performed in colon sections from parafibromin cKO mice. Scale bars, 50 μm. (i,j) Parafibromin cKO results in demise of intestinal organoids. Time-lapse image (i) and number (j) of intestinal organoids established from control and parafibromin cKO mice at each day after 4-OHT treatment (n=3, mean±s.d. ^**P<0.01; a two-tailed unpaired Student's t-test). Scale bars, 100 μm. (k–m) BMP signalling regulates PTK6 expression. Intestinal organoids were treated with recombinant BMP4 and subjected to immunoblotting (k,l) or CD44 immunostaining (m). (n) Parafibromin is essential for the development of the intestinal epithelium. H&E staining of embryonic intestinal tissues from control and Hrpt2^flox/flox/Villin-Cre embryos at E16.5. Scale bars, 50 μm.