1. Volume 115, Issue 2, Pages 297-306 (August 1998)
Cyclooxygenase 2 is induced in colonic epithelial cells in inflammatory bowel disease 
Irwin I. Singer*, Douglas W. Kawka*, Suzanne Schloemann‡, Teresa Tessner‡, Terrence Riehl‡, William F. Stenson‡ 
Gastroenterology 
Volume 115, Issue 2, Pages (August 1998)
DOI: /S (98)
Copyright © 1998 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Detection of COX-1 and COX-2 in epithelial cells isolated from normal, Crohn's colitis, and ulcerative colitis mucosa by Western blotting. Lysates were made of epithelial cells isolated from Crohn's colitis, ulcerative colitis (UC), and normal colonic mucosa. Sheep COX-1, sheep COX-2, lysates of unstimulated HUVECs, and lysates of HUVECs incubated with LPS and TNF-α were included as controls. Samples were separated by 8% sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Immunoblotting was performed using (A) polyclonal rabbit anti-sheep COX-1 antibody no. 241 and (B) polyclonal rabbit anti-sheep COX-2 antibody no Bands were detected by enhanced chemiluminescence.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Detection of COX-2 mRNA in normal colonic mucosa and ulcerative colitis mucosa by RPAs. Human COX-2 and GAPDH mRNA levels were assayed by RPA. In vitro transcription was performed to produce 32P-labeled antisense probes: 400 base pairs for COX-2 and 434 base pairs for GAPDH. Total RNA (20 μg) extracted from the mucosa of human colonic surgical resections was hybridized with excess 32P probe as described in Materials and Methods. mRNA was quantified by comparing the densitometric intensity of the protected bands from the samples with the protected bands from known amounts of in vitro transcribed sense-strand standards. This figure is a representative COX-2 RPA with samples from a normal colon, an ulcerative colitis colon, and three different quantities of sense-strand control RNA.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

4. Fig. 3
COX-1 is expressed (A) in crypt epithelial cells in Crohn's ileitis and (B) in epithelial cells in the lower half of the crypt in normal human colonic mucosa. Immunohistochemical localization of COX-1 colon was assessed in tissue fixed in Bouin's solution and embedded in paraffin. Immunoreactive COX-1 was detected by incubation with rabbit anti-mouse COX-1 antibody at a 1:2000 dilution. Bound antibody was visualized using the fluorescein-conjugated tyramide signal amplification system (green fluorescence). (A) In Crohn's ileitis, COX-1 is localized to the crypt epithelial cells (closed arrow) and to lamina propria mononuclear cells (arrowhead) but not to villus epithelial cells (open arrow). (B) In normal human colon, COX-1 is localized to crypt epithelial cells in the bottom half of the crypt (closed arrow) and to scattered lamina propria mononuclear cells (arrowhead) but not to epithelial cells in the upper half of the crypt (open arrow) (bar = 100 μmol/L).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Immunolocalization of COX-2 expression in the normal and ulcerative colitis colon. Adjacent sections of (A and B) normal or (C–F) colitic human colon were stained (A and C) with H&E, (B and D) with Dina IgG recognizing COX-2, or (E and F) with no. 168 IgG recognizing COX-2. (A) Histologically normal colon shows an intact epithelium and no evidence of inflammation. (B) No COX-2 expression is detected in the apical (arrowheads) or crypt (arrows) epithelium or within the lamina propria (*). (C) Ulcerative colitis colon shows crypt dilatation (arrows) and inflammation and edema within the lamina propria. (D) Intense expression of COX-2 can be seen in the apical epithelium (arrowheads) and in cells of the lamina propria (*) but not within the crypt epithelium (arrows). (E) High levels of COX-2 labeling are detected in the surface epithelium in ulcerative colitis (arrowheads) by affinity-purified no. 168 IgG in the presence of control COX-1 peptide. (F) Incubation with the COX-2–blocking peptide completely prevents staining of the surface epithelium (arrowheads) with the no. 168 anti–COX-2 IgG (bar = 100 μmol/L).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Localization of COX-2 in (A) Crohn's ileitis and (B) Crohn's colitis. (A) In Crohn's ileitis, COX-2 is expressed in villus epithelial cells (arrow) and lamina propria mononuclear cells (arrowhead) but not in crypt epithelial cells. In the villus epithelial cells, COX-2 is expressed in the perinuclear area, especially at the base of the nucleus, and in the Golgi. (B) In Crohn's colitis, COX-2 is expressed in surface epithelial cells (arrow) and scattered lamina propria mononuclear cells (arrowhead) but not in crypt epithelial cells. In the surface epithelial cells, COX-2 is expressed in the perinuclear area, especially at the base of the nucleus.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Expression of COX-2 and iNOS are colocalized in the surface epithelium of the ulcerative colitis colon. (A) H&E-stained section showing crypt dilatation and inflammation of the lamina propria. (B) Adjacent section stained with Dina anti–COX-2 IgG shows intense COX-2 localization in the surface epithelium (arrowheads) that abruptly diminishes at the zone of transition into the proximal portion of the crypt (arrows). (C) Neighboring section shows intense iNOS expression localized in the same regions of the colonic epithelium that showed high COX-2 labeling (corresponding arrowheads and arrows in B). (D) Another section stained with nonimmune rabbit IgG is completely unlabeled (bar = 100 μmol/L).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions