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brom bones possesses an increased level of Notch signaling activity in the intestinal epithelium. (A–D) Cross-sections of the anterior intestines from control (A,C) and mutant fish (B,D) immunostained for Hes1 (A,B) and NICD (C,D). Arrowheads in A and B point to Hes1 positive-cells. Arrows in (C) and (D) indicate NICD-positive cells. (E) Quantification of the NICD-positive cell number. Values shown are the percentages of NICD-positive cells relative to the total number of intestinal epithelial cells per villous cross-section (n = 114 villi from 7 brom bones heterozygous fish and 175 villi from 10 brom bones homozygous fish). Error bars represent s.e.m; P = 0.001 by Student's t test. (F,G) Real-time RT-PCR results show the expression levels of her6 (F) and her9 (G) in the intestines of 4 wild-type fish and 4 brom bones homozygotes. The expression levels of her6 and her9 were normalized to odc. Gene expression levels in the wild-type 1 fish were set as 100%. Scale bar = 50 μm.
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