Xenbase Image ID: 154230

Figure 6. Inhibition of Notch signaling by dibenzazepine (DBZ) leads to decreased cell proliferation and secretory cell hyperplasia. The intestine isolated from stage-56/57 tadpoles was cultured with 20 nM T3 for 5 (A-F) or 7 (G-J) days in the presence of DMSO (A, C, E, G, I) or 10 μM DBZ (B, D, F, H, J) after precultivation of the isolated intestinal fragments with DMSO or 10 μM DBZ for 3 days (see Fig. 4A). Cryosections of the intestinal fragments were hybridized with antisense Hairy1 (A, B) or Hairy2b (C, D). Paraffin sections were incubated with anti-PCNA (E, F) or anti-IFABP (G, H) antibodies or subjected to Periodic acid-Schiff (PAS) staining followed by counter-staining with hematoxylin (I, J). As a control, paraffin sections of the intestine from wild type froglets at stage 66 were also stained with PAS to identify goblet cells (K, arrowheads). Note that after 5 days, TH-induced up-regulation of Hairy genes (A, C, arrowheads) was impaired by DBZ treatment in vitro (B, D). PCNA-positive epithelial cells also decreased by DBZ treatment (E, F, arrowheads). After 7 days, IFABP was detected in most of the newly formed adult epithelial cells of the control intestine (G, arrowheads), but not the DBZ-treated intestine (H). A small number of goblet cell was detected in the control intestine (I, arrowhead), whereas PAS-positive secretory cells increased by DBZ treatment (J, arrowheads). The dashed-lines indicate the boundary of the epithelium and the connective tissue. Scale bars = 20 μm. Abbreviations: CT, connective tissue, DBZ, dibenzazepine, Ep, epithelium, PAS, Periodic acid-Schiff Image published in: Hasebe T et al. (2017) © 2016 The Authors STEM CELLS published by Wiley Periodicals, Inc. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives license Larger Image Printer Friendly View

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