goat polyclonal anti st2 antibody  (R&D Systems)

Journal: Cancer Immunology Research

Article Title: Interleukin 33 Signaling Restrains Sporadic Colon Cancer in an Interferon-γ–Dependent Manner

doi: 10.1158/2326-6066.cir-17-0218

Figure Lengend Snippet: Figure 1. St2 deficiency promotes the development of sporadic colon cancer. Sporadic colon cancer was induced by weekly administration of AOM over 6 consecutive weeks (6AOM) to WT and St2/ mice and analysis occurred 9–15 weeks after the last AOM injection depending on the onset of tumor development. A, Schematic of the experimental procedures. B, Representative whole mounts of the colons of WT and St2/ mice following 6AOM treatment 15 weeks after last AOM injection (left), and enumeration of the total number of resulting tumors per mouse (right). C and D, Representative sections from formalin-fixed and paraffin- embedded colons stained for hematoxilin–eosin (C; scale bar, 100 mm) and Ki67 or Apoptag (D; scale bar, 25 mm). E, Enumeration of positive staining cells from D expressed as the total number of Ki67þ cells (left) or Apoptagþ cells (right) per mm2 of tumor area. F and G, Quantitative RT-PCR analysis for Il33 (F) and Il1rl1 (exons 10–11, specific for membrane-bound isoform) expression (G) from tumors or adjacent normal colon tissues from 6AOM-treated WT and St2/ mice. Data were normalized against expression of Gapdh. B–G, Data were pooled from three independent experiments and shown as mean SEM, with each symbol representing an individual mouse. Data were analyzed using unpaired Student t test (E) or unpaired t test with Welsh correction (B, F). , P < 0.01; , P < 0.05; n.s., not significant.

Article Snippet: The sections were blocked with TNB Buffer (0.1 mol/L Tris–HCl, 0.15 mol/L NaCl, and 0.5% Blocking Reagent; PerkinElmer, Inc.) prior to staining with polyclonal goat anti-mouse St2 (R&D Systems, catalog # AF1004) and polyclonal goat anti-mouse IL33 (R&D Systems, catalog # AF3626).

Techniques: Injection, Staining, Quantitative RT-PCR, Membrane, Expressing

Journal: Cancer Immunology Research

Article Title: Interleukin 33 Signaling Restrains Sporadic Colon Cancer in an Interferon-γ–Dependent Manner

doi: 10.1158/2326-6066.cir-17-0218

Figure Lengend Snippet: Figure 3. IL33 stimulation induces expression of NF-kB target genes in colon-derived mesenchymal cells. A, Quantitative RT-PCR analysis of Il1rl1 mRNA expression encoding all isoforms (exons 7–8) or membrane-bound isoform (exons 10–11) in EpCAMþ/CD45 epithelial cells and EpCAM/CD45 mesenchymal cells isolated from unaffected colon ("colon") and tumors ("tumor") of WT mice undergoing the 6AOM protocol or left untreated ("UT"). Data were derived from 2 to 4 individual mice per cohort and were normalized against expression of Hprt. Data are representative of two (untreated colon) or one (colon and 6AOM colon and tumor samples) independent experiments. Mean SEM of the technical replicates are shown. B, Mesenchymal cells (CD45/EpCAM/CD90þ FSCHi) were isolated from the colon of WT mice and cultured in vitro for 16 hours. FACS plots show St2 and CD90 expression after 16 hours in vitro culture and is representative of three independents experiments. C and D, Cytokine production by mesenchymal cells, isolated as in B, and stimulated in vitro for 16 hours with IL33 (20 ng/mL) or left untreated. Quantitative RT-PCR expression analysis of indicated genes relative to expression of Hprt (C). Protein quantification following multiplex cytokine assay in the supernatant of cultured mesenchymal cells in D. Mean SEM with symbols representing independent experiments. Data were analyzed using paired Student t test. , P < 0.001; , P < 0.01; , P < 0.05. E, Immunobloting analysis of NIH-3T3 murine embryonic fibroblasts stimulated with murine IL33 (20 ng/mL).

Article Snippet: The sections were blocked with TNB Buffer (0.1 mol/L Tris–HCl, 0.15 mol/L NaCl, and 0.5% Blocking Reagent; PerkinElmer, Inc.) prior to staining with polyclonal goat anti-mouse St2 (R&D Systems, catalog # AF1004) and polyclonal goat anti-mouse IL33 (R&D Systems, catalog # AF3626).

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Membrane, Isolation, Cell Culture, In Vitro, Multiplex Assay, Cytokine Assay, Western Blot

Journal: Cancer Immunology Research

Article Title: Interleukin 33 Signaling Restrains Sporadic Colon Cancer in an Interferon-γ–Dependent Manner

doi: 10.1158/2326-6066.cir-17-0218

Figure Lengend Snippet: Figure 4. Loss of IL33 signaling in both the radiosensitive and the radioresistant cell compartments promotes Treg infiltration within colon tumors. A–C, Flow cytometric analysis of enzymatically digested tumors collected from 6AOM-treated BM chimeric mice generated as in Fig. 2. Representative FACS plots show St2 and Foxp3 expression on gated CD4þ/CD3þ/CD90High/CD45þ/EpCAM T cells (A). Data show Foxp3þ/CD4þ Treg among CD45þ/EpCAM hematopoietic cells (B, left), and ratio of Treg to EpCAMþ/CD45 tumor cells (B, right). Frequency of St2þ cells among Foxp3þ/CD4þ Tregs within the spleen or tumors from the indicated BM chimeras (C). A–C, Bars show mean SEM; symbols represent individual mice. D–G, Flow cytometric analysis of enzymatically digested colon tumors from 6AOM-treated WT or St2/ mice. Representative FACS plots showing intracellular expression of Gata3 and Rorgt in Foxp3þ/CD4þ Tregs (D). Frequency of Gata3þ

Article Snippet: The sections were blocked with TNB Buffer (0.1 mol/L Tris–HCl, 0.15 mol/L NaCl, and 0.5% Blocking Reagent; PerkinElmer, Inc.) prior to staining with polyclonal goat anti-mouse St2 (R&D Systems, catalog # AF1004) and polyclonal goat anti-mouse IL33 (R&D Systems, catalog # AF3626).

Techniques: Generated, Expressing

Journal: Cancer Immunology Research

Article Title: Interleukin 33 Signaling Restrains Sporadic Colon Cancer in an Interferon-γ–Dependent Manner

doi: 10.1158/2326-6066.cir-17-0218

Figure Lengend Snippet: Figure 5. Loss of IL33 signaling in the radioresistant cell compartment leads to a decrease in IFN gene signature and immune activation within colon tumors. Whole genome expression analysis of tumors collected from BM chimeras generated as in Fig. 2. A, Distribution of commonly and uniquely differentially expressed genes between tumors collected from the indicated BM chimeras (red—upregulated, blue—downregulated). B, Relative expression level (z score) of the 65 DE genes commonly regulated in BM chimeras in which St2 deficiency was restricted to the radioresistant compartment as indicated in A. C, Metacore analysis of biological processes for the 65 DE genes selected as in B. D, Normalized expression counts of selected genes in tumors from the indicated BM chimeras as determined by RNAseq. Mean SEM normalized expression counts with each symbol representing an individual mouse.

Article Snippet: The sections were blocked with TNB Buffer (0.1 mol/L Tris–HCl, 0.15 mol/L NaCl, and 0.5% Blocking Reagent; PerkinElmer, Inc.) prior to staining with polyclonal goat anti-mouse St2 (R&D Systems, catalog # AF1004) and polyclonal goat anti-mouse IL33 (R&D Systems, catalog # AF3626).

Techniques: Activation Assay, Expressing, Generated