Fig 1: 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.
Fig 2: Dectin-3-/- mice have increased tumor load upon AOM-DSS treatment than WT miceSingle-housed WT mice and Dectin-3-/- mice (n = 8 for each group) were injected intraperitoneally with one dose of AOM (10 mg/kg), followed by three cycles of feeding water with 2% DSS. After induction of tumorigenesis, mice were euthanized on day 100. ARepresentative images of colon tumors were shown. Scale bars, 10 mm.BTumor number, tumor size, and tumor load in each mouse were measured.C, DHistological analysis of colon tumors was shown by hematoxylin and eosin (HE) staining. Tumors were microscopically analyzed and classified as low or high grade. Histological score was assessed by a pathologist. Scale bars, 25 µm.E, FTumor tissues were stained for Ki67 and p-STAT3. The percentage of Ki67-positive and p-STAT3-positive cells was quantified. Scale bars, 25 µm.GProportion of CD11b+F4/80+ and ROR?t+/CD45+lin- cells was analyzed in colonic LP cells and mLNs among tumor-bearing WT and Dectin-3-/- mice by flow cytometry.HRelative expression of Il-6, Il-22, Cxcl1, and Il-17a in LP cells from tumor-bearing WT and Dectin-3-/- mice were detected using qPCR. 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. See also Fig EV1.
Fig 3: Up-regulation of IL-22 in Dectin-3 -/- mice contributes to CAC development A, BMice were treated as described in Fig 2A. LP cells were isolated from each mouse and were culture for 48 h. Cytokine and chemokine production of LP cells were detected using multiplex cytokine assay. Color from blue to red indicates enrichment of gene expression.CmRNA expressions of IL-22, ß-defensin, Reg3g, and Cxcl1 in LP cells were detected by qPCR.DLP cells and mLNs were isolated from each mouse and were culture for 48 h. Production of IL-22 in LP cells and mLNs were detected by ELISA.ELP cells were isolated from each mouse and were culture for 48 h. Expression of Il-17 in LP cells were detected by qPCR. Production of IL-17 in LP cells were detected by ELISA.FProtein level of p-STAT3 and STAT3 in colonic epithelial cells was detected by using Western blot (one mouse per lane). The relative expression of p-STAT3 was calculated.GWT and Dectin-3-/- mice were intraperitoneally treated with anti-IL22 antibody or anti-IgG antibody as control during AOM-DSS administration (n = 5, each group). Mice were euthanized on Day 100, tumor number, tumor size, and tumor load in colons were measured. 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. See also Fig EV3.
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