1. Volume 125, Issue 2, Pages 477-490 (August 2003)
Identification of a quantitative trait locus for ileitis in a spontaneous mouse model of Crohn’s disease: SAMP1/YitFc 
Kosuke Kozaiwa, Kazuhiko Sugawara, Michael F Smith, Virginia Carl, Vladimir Yamschikov, Brian Belyea, Sherri B Mcewen, Christopher A Moskaluk, Theresa T Pizarro, Fabio Cominelli, Marcia Mcduffie 
Gastroenterology 
Volume 125, Issue 2, Pages (August 2003)
DOI: /S (03)00876-X

2. Figure 1
Allele determination shows a unique set of non-AKR alleles in SAMP1/Fc mice. Microsatellite analysis using an automated sequencer shows the complexity of the SAMP1/Fc genetic background. Recombination distances in cM/L were obtained from the Mouse Genome Database (The Jackson Laboratories; and the relative positions were confirmed by analysis of B6SF2 and B6S backcross mice (data not shown). Allele sizes found in AKR mice are marked as dark gray squares, with light gray squares designating a shared fragment size with other strains in the test panel. Black squares show non-AKR allele sizes shared with other strains in the test panel. Absence of fill across the panel for any microsatellite locus indicates that the fragment size from SAMP1/Fc is unique among the tested strains. The comparison strains include mice from 6 major lineages and represent most of the common gene pools among laboratory mouse strains.
Gastroenterology  , DOI: ( /S (03)00876-X)

3. Figure 2
Distribution of total inflammatory scores in SAMP1/Fc, B6SF1, and B6SF2 mice. Active inflammatory index (severity and extent of granulocytic infiltration), chronic inflammatory index (severity and extent of mononuclear cell infiltration), and an index of villus architectural changes were scored on the terminal 15 cm of small intestine from mice aged 30 weeks as described in Materials and Methods. The 3 indices were added to give a total inflammatory score.
Gastroenterology  , DOI: ( /S (03)00876-X)

4. Figure 3
Evidence for linkage of the total inflammatory score to Chr 6, 9, 17, and X. Single-point LRS scores were calculated for a panel of microsatellites sampling the entire mouse genome (except for the Y chromosome) using Map Manager QT. (A) LRS scores >9.5 (P < 0.01) were detected on Chr 9, 17, and X in the initial analysis using selected cohorts representing extreme phenotypes (solid line). After stratification by genotype at the locus representing peak linkage on Chr 9 (D9Mit123), additional evidence suggestive of linkage was detected on Chr 6 (gray fill). (B) The entire F2 cohort (n = 150) was genotyped at increased marker density on all chromosomes with LRS scores >7.0, including loci from Chr 1, 2, 8, and 19 (data not shown). Evidence of linkage persisted on Chr 9, with the highest LRS values obtained for new loci in the region centromeric to D9Mit123, and on Chr X (solid line). Once again, LRS values increased on Chr 6 following stratification based on genotype at D9Mit191 (gray fill). Further stratification (for loci on Chr 6 or X) did not show additional evidence of linkage across any of the intervals genotyped in the entire F2 cohort. No evidence of additional distant loci on Chr 9 was detected in either experiment following stratification by the locus with peak evidence for linkage. (C) B6SF2 mice were grouped according to haplotype from D9Mit297 to D9Mit308: S/S, SAMP homozygotes; B/S, presumed heterozygotes; B/B, B6 homozygotes; Rec, defined recombinants. Total inflammatory scores are shown for each mouse, with boxes enclosing the second and third quartiles. Comparison of scores in each group using an unpaired t test suggested a dose-dependent effect for the Chr 9–linked susceptibility locus in this cross.
Gastroenterology  , DOI: ( /S (03)00876-X)

5. Figure 4
Comparative mouse/human map of the SAMP1/Fc susceptibility locus on Chr 9. Interval mapping using genotype results from the entire B6SF2 cohort suggested peak linkage between D9Mit191 and D9Mit48 (black bar). Evidence of linkage at significant (gray bars) and suggestive (hatched bars) levels was determined by permutation analysis using all genotypes on Chr 9. Recombination distances were calculated using Map Manager QT, and relative distances and order were confirmed using version 3 of the Mouse Genome Assembly for all currently mapped loci. Potential candidate genes were identified based on known expression levels in the immune system or the intestine. Genes expressed in lymphoid or myeloid lineages are in italics, and genes expressed in intestinal epithelium are in bold.
Gastroenterology  , DOI: ( /S (03)00876-X)

6. Figure 5
IL-10 receptor α chain polymorphisms and function. (A) Comparative genomic sequence for the IL-10 receptor α locus was performed as described. Two single-nucleotide insertions in introns 1 and 3 distinguish B6 from AKR or SAMP1/Fc. (B) Cell lysates from macrophages cultured with or without 5 ng/mL human IL-10 were harvested at the designated time points after addition of human IL-10. Electrophoresis was followed by sequential immunoblotting with anti-phosphorylated Stat3-Tyr705 and anti-Stat3 antibodies. (C) Tumor necrosis factor α messenger RNA levels were quantified in total RNA from bone marrow-derived macrophages from SAMP1/Fc (filled bars) and B6 mice (open bars) after stimulation for 3 hours with 100 ng/mL lipopolysaccharide. Tumor necrosis factor α messenger RNA was below the limits of detection in unstimulated control cells or in cells exposed to IL-10 alone.
Gastroenterology  , DOI: ( /S (03)00876-X)

7. Figure 6
IL-18 protein levels in SAMP1/Fc mice. A significant increase in circulating IL-18 serum levels was observed in SAMP1/Fc mice at 4 weeks of age compared with age-matched AKR mice (P < 0.05); although a comparable trend was also found between SAMP1/Fc with B6 mice, this observed difference did not reach statistical significance (P < 0.08). Increased expression of IL-18 protein was also found in the MLNs of 4-week-old SAMP1/Fc mice relative to both AKR and B6 tissues (P < 0.05), suggesting that up-regulation of IL-18 occurs at a very early stage in the disease process in SAMP1/Fc mice.
Gastroenterology  , DOI: ( /S (03)00876-X)