1. Volume 122, Issue 2, Pages 469-482 (February 2002)
Disruption of hedgehog signaling reveals a novel role in intestinal morphogenesis and intestinal-specific lipid metabolism in mice 
Li Chun Wang, Fatiha Nassir, Zhong–Ying Liu, Leona Ling, Frank Kuo, Thomas Crowell, Dian Olson, Nicholas O. Davidson, Linda C. Burkly 
Gastroenterology 
Volume 122, Issue 2, Pages (February 2002)
DOI: /gast
Copyright © 2002 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Expression of hh and Ptc-1 protein during intestinal development. Protein extracts (A) from gut tissues of E15, E17, and E19 mice and mice of postnatal day 2 and 3 (P2 and P3), (B) from duodenum (D), jejunum (J), ileum (I), and cecum (Ce) of 2-week-old mice (C) from intestinal epithelial cell lines T84 (human), IEC6 (rat), Caco-2 (human), SW480 (human), and from intestinal fibroblast cell line CCD18 (human) were subjected to immunoprecipitation-Western blot analysis using control or anti-hh mAb. (C) Control mAb is indicated as C and anti-hh mAb is indicated as +. Anti-hh mAb specifically detected the 19-kilodalton hh protein. Expression of Ptc-1 was evaluated in Ptc1-LacZ negative (Ptc−) and Ptc1-LacZ positive (Ptc+) mice. Ptc-1 expression was visualized by x-gal staining of gut tissues from (D) E15.5 embryos, (E) P1, or (F) adult small intestines. Note that the blue/purple x-gal staining displayed along the surface of the villi in (E) were frequently observed in the postnatal intestinal tissues of both the Ptc− and Ptc+ mice. Scale bars: (D) 100 μm; (E) 50 μm; and (F) 200 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Postnatal lethality and vacuolation of enterocytes in anti-hh mAb–treated mice. Mice at postnatal day 3 (A, left) and 6 (A, middle) were treated with anti-hh or control mAb in utero from E12.5 (E12.5/P3 and E12.5/P6). Mice at postnatal day 14 (A, right) were treated with anti-hh or control mAb from the day of birth (P1/P14). The 2 mice on the left in each figure were treated with anti-hh mAb, whereas the 2 mice on the right were treated with control mAb. (B) Intestinal tissues from mice at P1 or P2 from mice treated with anti-hh or control mAb in utero starting at E12.5. (C) Intestinal tissues from mice at P17 from mice treated with anti-hh or control mAb starting at P2. Tissues were stained with H&E. Scale bars: (B and C) 50 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Intestinal epithelial cell migration, lineage differentiation, and lipid malabsorption in anti-hh mAb–treated mice. (A) Mice were treated with control mAb (left) or anti-hh mAb (right) starting at P2 and killed at P15 (pulse) or P18 (chase) after pulse-chase labeling with BrdU. (B) Paraffin-embedded small intestinal tissues were subjected to histochemistry analyses using anti-BrdU mAb, and PAS–Alcian-blue staining of goblet cells was performed at P15. (C) For Oil Red O staining, mice were treated with either control (top) or anti-hh mAb (bottom) starting at P1 and killed at P13. H&E (left) or Oil Red O staining of lipid (right) was performed on small and large intestinal tissues. Shown here are representative ileum tissues. (D) Fecal fat microscopy was performed from fecal samples of P13 mice treated with control (left) or anti-hh mAb (right) starting at P2. Scale bars: (A–C) 50 μm; (D) 100 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Apolipoprotein and lipoprotein profiles in anti-hh mAb–treated mice. Mice were treated with either control or anti-hh mAb starting at P1; sera were collected at P10, P12, and P15; and (A) Western blot was performed using anti-apoA-IV antibody or (B) lipoprotein profile was performed on serum at P15. (▵(, anti-hh mAb; ■, control mAb). Results are presented as arbitrary units. The elution peak for each lipoprotein class is indicated in (B).
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Expression of MTP and apo B mRNAs in the intestinal tissues of the anti-hh mAb–treated mice. Mice were treated with anti-hh mAb (n = 3, left) or control mAb (n = 3, right) starting at P1 and killed at P12. Total RNA was hybridized with (A) 32P-labeled MTP or (B) apoB RPA probe and subjected to RPA analysis followed by autoradiography. The mRNA level of the corresponding housekeeping gene, GAPDH, was included to normalize for gel loading. (C) ApoB mRNA editing was measured by primer extension analysis as described in Materials and Methods and subjected to phospho-Imager analysis. U, uridine; C, cytidine; P, primer; %U, percentage of C to U deamination. Controls for edited UAA (U, second right lane) and unedited CAA mRNA (C, first right lane) consist of in vitro editing of an apoB template.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Large cytosolic lipid droplets in the enterocytes of anti-hh mAb–treated mice. Mice were treated with either control or anti-hh mAb starting at P1; at P13, the intestinal (ileum) tissue was subjected to electronic microscopic analysis. Note that the height of microvilli indicated by the arrow was reduced in anti-hh–treated mice. *Large cytosolic lipid droplets and **mucin present in the goblet cells. Scale bars: 2.5 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

8. Fig. 7
Disruption of hh signaling by anti-hh mAb treatment. (A) E14.5 pregnant mice were treated with control or anti-hh mAb and the in vivo binding activity of the mAbs was examined in E15.5 intestines. The anti-hh mAb bound specifically to epithelial layer of the intestine. (B) Expression of Ptc-1 was examined in the intestinal tissues of Ptc+ mice treated with 20 μg of control or anti-hh mAb 3 times a week starting at P8. Mice were killed at P19. Representative jejunum tissues are shown. The net expression of Ptc-1 was reduced in anti-hh mAb treated intestines. Scale bars: (A) 200 μm; (B) 100 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions