MOLF express high levels of CHI3L1 in the colon and polypoid area and associates with high immune cellular infiltrates.

<p>Two-color Immunofluorescence analysis was performed on colonic sections from six months old B6 and MOLF mice using anti-CHI3L1 and anti-F4/80 (<b>A</b>) -CD11b (<b>B</b>) or -CD4 (<b>C</b>) primary antibodies respectively.</p

MOLF express high levels of CHI3L1 in colonic epithelial cells but not in immune cells.

<p>Immunohistochemical analysis was performed on colonic serial sections (non-polypoid area) from six months old MOLF mice using anti-CHI3L1, -TROMA–1 (cytokeratin 8), -CD4, -CD11b, and -F4/80 antibodies.</p

MOLF mice spontaneously developed polyp-like nodules in colon at steady state.

<p>[<b>A</b>] Six months old MOLF and B6 mice were sacrificed and the colons were opened up longitudinally. [<b>B</b>] Multiplicity and size of polyp-like nodules were determined under a dissection microscope (n = 7 in each group). [<b>C</b>] H&E sections of colons from B6 and MOLF are shown. [<b>D</b>] IHC staining of colonic sections from B6 and MOLF mice was performed using anti-Ki67 primary antibody.</p

High endogenous colonic CHI3L1 expression state MOLF/EiJ mice colon at steady state.

<p>[<b>A</b>] Colonic CHI3L1 mRNA level in MOLF/Eij (MOLF) and C57BL/6 (B6) mice were measured using real time PCR (n = 5 in each group). [<b>B</b>] Serum CHI3L1 protein level in MOLF (n = 5) and B6 (n = 5) mice were quantified using ELISA. [<b>C</b>] Schema showing the genomic structure of the CHI3L1 locus. Arrows indicating single nucleotide polymorphisms between MOLF and B6 mice identified by in silico comparison using the Jackson mouse phenome database. [<b>D</b>] Proximal promoter from MOLF and B6 mice was PCR amplified, sequenced and aligned using Clustal W program. [<b>E</b>] SW480 cells were transfected with pGL3-basic vector containing MOLF- or B6-derived CHI3L1 proximal promoter and renilla-normalised luciferase assay was performed (n = 5 in each group). **P<0.01, ***P<0.001 between the two groups as indicated.</p

Alignment of Human, MOLF and B6 mice CHI3L1 proximal promoter.

<p>Approximately 600 bp upstream sequence of the CHI3L1 transcription start site from human, MOLF and B6 mice were aligned using Clustal W program.</p