1. Volume 114, Issue 5, Pages 965-974 (May 1998)
Regional differences in L-selectin expression in murine intestinal lymphocytes 
Frank Seibold*, Beatrice Seibold-Schmid‡, Yingzi Cong*, Feng Yu Shu§, Robert P. McCabe*, Casey Weaver‡, Charles O. Elson* 
Gastroenterology 
Volume 114, Issue 5, Pages (May 1998)
DOI: /S (98)
Copyright © 1998 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Proportion of L-selectin–positive CD4+ T cells in the colon in the presence or absence of colitis. Colon IELs were isolated from groups of mice with no, mild, or severe colitis as described in Materials and Methods. Cells were stained with both anti–L-selectin and anti-CD4+ and then were analyzed by two-color flow cytometry, gating on the CD4+ population. None, healthy C3H/HeJ mice; mild colitis, spontaneously colitic C3H/HeJBir mice; and severe colitis, C3H/SnJ scid/scid mice that had received C3H/HeJ CD4+ CD45RBhi T cells 10 weeks before analysis. These data represent the means ± SD of 4–5 animals in each group from 2 separate experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Proportion of L-selectin–positive small intestinal IELs cultured in vitro for various times. After culture, cells were stained with anti–L-selectin and analyzed by flow cytometry. An aliquot incubated for 48 hours was exposed to 0.1 μmol/L PMA for 30 minutes before staining and analysis (48 hours +PMA). These data represent the means ± SD of replicates from 1 of 3 experiments with similar results.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Reverse-transcription PCR for L-selectin mRNA. Total RNA was isolated and reverse transcribed to cDNA, and L-selectin mRNA was amplified by PCR using specific primers. The products were electrophoresed on agarose gels and stained with ethidium bromide. Lane 1, small intestinal IELs; lane 2, colonic IELs; lane 3, spleen cells; lane 4, Mode K epithelial cell line; and lane 5, liver homogenate. Size markers are shown to the left of lane 1. This experiment was repeated three times with similar results.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

5. Fig. 4
L-Selectin expression on CD4+ MLN incubated in vitro with various stimulants. (A) Untreated MLN; (B) MLN incubated with 0.1 μmol/L PMA for 30 minutes; (C) MLN incubated with homogenized small intestine for 2 hours; and (D) MLN incubated with colon homogenate for 2 hours. The data are representative of the results of 3 independent experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

6. Fig. 4
L-Selectin expression on CD4+ MLN incubated in vitro with various stimulants. (A) Untreated MLN; (B) MLN incubated with 0.1 μmol/L PMA for 30 minutes; (C) MLN incubated with homogenized small intestine for 2 hours; and (D) MLN incubated with colon homogenate for 2 hours. The data are representative of the results of 3 independent experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

7. Fig. 5
Inhibition of L-selectin down-regulation by a metalloproteinase inhibitor. L-Selectin expression was measured on spleen cells incubated in vitro under various conditions that were then stained with anti–L-selectin antibody and analyzed by flow cytometry. (A) untreated control; (B) cells incubated with 0.1 μmol/L PMA for 30 minutes; (C) the same as in B but in the presence of metalloproteinase inhibitor KD-IX-73-5; (D) cells incubated with small intestinal homogenate; and (E) the same as in D but in the presence of metalloproteinase inhibitor KD-IX The data are representative of results of 3 independent experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

8. Fig. 5
Inhibition of L-selectin down-regulation by a metalloproteinase inhibitor. L-Selectin expression was measured on spleen cells incubated in vitro under various conditions that were then stained with anti–L-selectin antibody and analyzed by flow cytometry. (A) untreated control; (B) cells incubated with 0.1 μmol/L PMA for 30 minutes; (C) the same as in B but in the presence of metalloproteinase inhibitor KD-IX-73-5; (D) cells incubated with small intestinal homogenate; and (E) the same as in D but in the presence of metalloproteinase inhibitor KD-IX The data are representative of results of 3 independent experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

9. Fig. 5
Inhibition of L-selectin down-regulation by a metalloproteinase inhibitor. L-Selectin expression was measured on spleen cells incubated in vitro under various conditions that were then stained with anti–L-selectin antibody and analyzed by flow cytometry. (A) untreated control; (B) cells incubated with 0.1 μmol/L PMA for 30 minutes; (C) the same as in B but in the presence of metalloproteinase inhibitor KD-IX-73-5; (D) cells incubated with small intestinal homogenate; and (E) the same as in D but in the presence of metalloproteinase inhibitor KD-IX The data are representative of results of 3 independent experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

10. Fig. 6
Loss of L-selectin is proportional to the amount of intestinal homogenate added to the lymphocytes. Lymphocytes (1 × 106) from MLN were incubated for 90 minutes with either small bowel (—2—) or large bowel (··◊··) homogenate at different concentrations (as shown). After incubation, lymphocytes were washed, stained with both anti– L-selectin and anti-CD4, and analyzed by flow cytometry. These data represent the means ± SD of 3 experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

11. Fig. 7
Competitive inhibition of L-selectin shedding by a metalloproteinase inhibitor. Lymphocytes (1 × 106) from MLN were preincubated with the metalloproteinase inhibitor KD-IX-73-5 for 15 minutes at various concentrations as shown. Then 300 μg/mL of either colon homogenate (··◊··) or small intestine (—2—) homogenate was added. After incubation for 90 minutes, lymphocytes were washed, stained with both anti–L-selectin and anti-CD4, and analyzed by flow cytometry. These data represent the means ± SD of 3 experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions