Fig 2: ELISA and immunohistochemistry confirm gene expression data in Fig. 6. Ldlr-/- mice were fed the four diets, and the levels of protein in jejunum were determined by ELISA or the number of goblet cells or Paneth cells in jejunum were determined by immunohistochemistry as described in Materials and methods. The gender, age, and number of mice per group for each panel are shown in supplemental Table S5. A: IL-36?. B: IL-23. C: IL-22. D: NOTCH2. E: DLL4. F: ATOH1. G: GFI1. H: Goblet cells/crypt. I: Goblet cells/villus axis. J: Paneth cells/crypt. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Fig 3: IL-22 is correlated with fungal burden in patients with colon cancer APatients with colon cancer were divided into four groups based on tumor stage. mRNA expressions of Dectin-3 in tumor tissues were detected by qPCR. The box represents the 25th–75th percentile, and the whisker plots represent the minimum and maximum percentiles.BPatients with colon cancer were divided into two groups based on fungal burden in the feces. mRNA expressions of Dectin-3 were compared between these two groups.COverall survival rate was shown by K-M survival curve.D, ETumor tissues were stained for IL-22 and p-STAT3. The percentages of IL-22-positive and p-STAT3-positive cells were quantified. mRNA expressions of IL22 and STAT3 were detected by qPCR. Scale bars, 25 µm. Data information: Data with error bars are represented as mean ± SD. Each panel is a representative experiment of at least three independent biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001 as determined by Student’s t-test.

Fig 4: Up-regulation of IL-22 in Dectin-3 -/- mice contributes to CAC development. Related to Fig 3 AGF mice were orally gavaged with C. albicans (twice a week, 1 × 107) during administration with AOM-DSS (n = 8, each group). After induction of tumorigenesis (100 days), mice were euthanized and colons were removed. Tumor number, tumor size, and tumor load in colons were measured. Total fungal burden of colon was detected by using qPCR. Clinical colitis scores were evaluated on day 56.BWT and Dectin-3 -/- mice were treated with fluconazole during administrated with AOM-DSS (n = 8, each group). Tumor number, tumor size, and tumor load in colons were measured. Total fungal burden of colon was detected by using qPCR.CWT and Dectin-3 -/- mice were treated with antibiotics during administrated with AOM-DSS (n = 8, each group). Tumor number, tumor size, and tumor load in colons were measured. Total fungal burden of colon was detected by using qPCR.D Dectin-3 -/- mice were oral transferred with feces (400 ml each time, twice a week) from tumor-bearing WT mice during administrated with AOM-DSS (n = 8, each group). After induction of tumorigenesis, mice were euthanized and colons were removed. Tumor number, tumor size, and tumor load in colons were measured.E, FMice were treated as described in Fig EV3D. Fungal ITS2 rDNA gene sequence was performed in each group. Principal component analyses (PCA) based on fungal composition and Shannon analysis related to alpha diversity were used. F: samples are from Dectin-3-/- mice, P: samples are from Dectin-3-/- mice treated with WT feces.GFungal-taxon-based analysis at the genus level in feces of mice. 1 Candidan 2 Fusarium 3 Kazachstania 4 Thermomyces 5 Phaeococcomyces 6 Mycosphaerella 7 Penicillium 8 Aspergillus 9 Coprinellus 10 Cryptococcus 11 Mortierella 12 Guehomyces 13 Preussia 14 Chaetomium.HELISA results for binding assays of Dectin-3 with a-mannans. Plates were coated with a-mannans and then were added with 100 ml/well recombinant Dectin-3 at indicated concentrations. Data information: Data with error bars are represented as mean ± SD. Each panel is a representative experiment of at least three independent biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001 as determined by Student’s t-test.

Fig 5: Dectin-3 -/- mice have increased tumor burden and impaired immune responses upon AOM-DSS treatment than WT mice. Related to Fig 1 Mice were treated as described in Fig 1A. AClinical colitis scores were evaluated on day 56.BTumor tissues were stained for cleaved-caspase 3. The percentages of positive cells were quantified.C, DColonic LP cells and mLNs were isolated from each mouse. The proportion of immune cells was determined by flow cytometry.E, FRelative expression of Il-6, Il-22, Cxcl1, TNF-a and Il-17 in mLNs cells and tumors from tumor-bearing WT and Dectin-3 -/- mice were detected using qPCR.GCytokine and chemokine production in the serum of WT and Dectin-3-/- tumor-bearing mice was determined by a multiple cytokine detection assay. Data information: Data with error bars are represented as mean ± SD. Each panel is a representative experiment of at least three independent biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001 as determined by unpaired Student’s t-test.