rmil 4  (PeproTech)

 
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    Name:
    Recombinant Murine IL 4
    Description:
    IL 4 is a pleiotropic cytokine that regulates diverse T and B cell responses including cell proliferation survival and gene expression Produced by mast cells T cells and bone marrow stromal cells IL 4 regulates the differentiation of naive CD4 T cells into helper Th2 cells characterized by their cytokine secretion profile that includes secretion of IL 4 IL 5 IL 6 IL 10 and IL 13 which favor a humoral immune response Another dominant function of IL 4 is the regulation of immunoglobulin class switching to the IgG1 and IgE isotypes Excessive IL 4 production by Th2 cells has been associated with elevated IgE production and allergy Recombinant murine IL 4 is a 13 5 kDa globular protein containing 121 amino acid residues
    Catalog Number:
    214-14-100UG
    Price:
    650.00
    Category:
    Recombinant Proteins
    Source:
    E.coli
    Reactivity:
    Rat Pig Intestinal Worm
    Purity:
    98.0
    Quantity:
    100UG
    Buy from Supplier


    Structured Review

    PeproTech rmil 4
    TET proteins facilitate CSR. ( A ) Flowchart of in vivo experiment. f.p., footpad. ( B ) Top: Flow cytometric analysis of CD19 + GL7 + Fas + GC B cells at the draining popliteal lymph nodes from WT and Tet2/3 DKO mice after NP-OVA immunization as in (A). Bottom: Decreased IgG1-switched cells among GC B cells in Tet2/3 DKO (YFP + GC B–gated) compared with WT mice (GC B–gated). ( C to F ) Quantification of experiments shown in (B). Data shown are aggregated results from two independent experiments. Means and SEs are shown. WT, n = 11; DKO, n = 12. ( G ) Flowchart of in vitro IgG1 switching. Cells were labeled with Cell trace violet and activated for 4 days with LPS and IL-4. ( H and I ) Flow cytometry plots (H) and quantification (I) of IgG1-switched B cells in Tet2/3 DKO ( n = 4) and WT ( n = 4) mice. Representative of at least three independent experiments. ( J ) Circular γ1 transcript was quantified by qRT-PCR and normalized to Gapdh and then to the level of WT. Representative experiment of two independent experiments with three technical replicates. ( K ) Flowchart of in vitro IgA switching. Cells were activated for 5 days with anti-CD40, <t>rmIL-4,</t> rmIL-5, and rhTGFβ. ( L to N ) Flow cytometry plots (L) and quantification (M and N) of IgG1-switched (M) and IgA-switched cells (N). Representative of three independent experiments with three technical replicates. Statistical significance was calculated using unpaired two-tailed t test. * P
    IL 4 is a pleiotropic cytokine that regulates diverse T and B cell responses including cell proliferation survival and gene expression Produced by mast cells T cells and bone marrow stromal cells IL 4 regulates the differentiation of naive CD4 T cells into helper Th2 cells characterized by their cytokine secretion profile that includes secretion of IL 4 IL 5 IL 6 IL 10 and IL 13 which favor a humoral immune response Another dominant function of IL 4 is the regulation of immunoglobulin class switching to the IgG1 and IgE isotypes Excessive IL 4 production by Th2 cells has been associated with elevated IgE production and allergy Recombinant murine IL 4 is a 13 5 kDa globular protein containing 121 amino acid residues
    https://www.bioz.com/result/rmil 4/product/PeproTech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rmil 4 - by Bioz Stars, 2021-05
    86/100 stars

    Images

    1) Product Images from "TET enzymes augment activation-induced deaminase (AID) expression via 5-hydroxymethylcytosine modifications at the Aicda superenhancer"

    Article Title: TET enzymes augment activation-induced deaminase (AID) expression via 5-hydroxymethylcytosine modifications at the Aicda superenhancer

    Journal: Science immunology

    doi: 10.1126/sciimmunol.aau7523

    TET proteins facilitate CSR. ( A ) Flowchart of in vivo experiment. f.p., footpad. ( B ) Top: Flow cytometric analysis of CD19 + GL7 + Fas + GC B cells at the draining popliteal lymph nodes from WT and Tet2/3 DKO mice after NP-OVA immunization as in (A). Bottom: Decreased IgG1-switched cells among GC B cells in Tet2/3 DKO (YFP + GC B–gated) compared with WT mice (GC B–gated). ( C to F ) Quantification of experiments shown in (B). Data shown are aggregated results from two independent experiments. Means and SEs are shown. WT, n = 11; DKO, n = 12. ( G ) Flowchart of in vitro IgG1 switching. Cells were labeled with Cell trace violet and activated for 4 days with LPS and IL-4. ( H and I ) Flow cytometry plots (H) and quantification (I) of IgG1-switched B cells in Tet2/3 DKO ( n = 4) and WT ( n = 4) mice. Representative of at least three independent experiments. ( J ) Circular γ1 transcript was quantified by qRT-PCR and normalized to Gapdh and then to the level of WT. Representative experiment of two independent experiments with three technical replicates. ( K ) Flowchart of in vitro IgA switching. Cells were activated for 5 days with anti-CD40, rmIL-4, rmIL-5, and rhTGFβ. ( L to N ) Flow cytometry plots (L) and quantification (M and N) of IgG1-switched (M) and IgA-switched cells (N). Representative of three independent experiments with three technical replicates. Statistical significance was calculated using unpaired two-tailed t test. * P
    Figure Legend Snippet: TET proteins facilitate CSR. ( A ) Flowchart of in vivo experiment. f.p., footpad. ( B ) Top: Flow cytometric analysis of CD19 + GL7 + Fas + GC B cells at the draining popliteal lymph nodes from WT and Tet2/3 DKO mice after NP-OVA immunization as in (A). Bottom: Decreased IgG1-switched cells among GC B cells in Tet2/3 DKO (YFP + GC B–gated) compared with WT mice (GC B–gated). ( C to F ) Quantification of experiments shown in (B). Data shown are aggregated results from two independent experiments. Means and SEs are shown. WT, n = 11; DKO, n = 12. ( G ) Flowchart of in vitro IgG1 switching. Cells were labeled with Cell trace violet and activated for 4 days with LPS and IL-4. ( H and I ) Flow cytometry plots (H) and quantification (I) of IgG1-switched B cells in Tet2/3 DKO ( n = 4) and WT ( n = 4) mice. Representative of at least three independent experiments. ( J ) Circular γ1 transcript was quantified by qRT-PCR and normalized to Gapdh and then to the level of WT. Representative experiment of two independent experiments with three technical replicates. ( K ) Flowchart of in vitro IgA switching. Cells were activated for 5 days with anti-CD40, rmIL-4, rmIL-5, and rhTGFβ. ( L to N ) Flow cytometry plots (L) and quantification (M and N) of IgG1-switched (M) and IgA-switched cells (N). Representative of three independent experiments with three technical replicates. Statistical significance was calculated using unpaired two-tailed t test. * P

    Techniques Used: In Vivo, Flow Cytometry, Mouse Assay, In Vitro, Labeling, Cytometry, Quantitative RT-PCR, Two Tailed Test

    2) Product Images from "The Anti-Inflammatory Immune Response in Early Trichinella spiralis Intestinal Infection Depends on Serine Protease Inhibitor–Mediated Alternative Activation of Macrophages"

    Article Title: The Anti-Inflammatory Immune Response in Early Trichinella spiralis Intestinal Infection Depends on Serine Protease Inhibitor–Mediated Alternative Activation of Macrophages

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.2000290

    The phenotypic effect of IL-4Rα blocking mAb on rmIL-4–treated or r Ts -serpin–treated peritoneal macrophages in vivo. ( A ) Blocking effect design of IL-4Rα blocking mAb in r Ts -serpin–treated or rmIL-4–treated peritoneal macrophages in vivo. Flow cytometry analysis of the F4/80 + ratio in nonlymphocyte subsets ( B ) and polarization in F4/80 + subsets ( C ) in rmIL-4–treated or r Ts -serpin–treated peritoneal cells coincubated with isotype control mAb or IL-4Rα blocking mAb. The values are the mean ± SD of eight independent experiments. Significant differences were analyzed by t test. ** p
    Figure Legend Snippet: The phenotypic effect of IL-4Rα blocking mAb on rmIL-4–treated or r Ts -serpin–treated peritoneal macrophages in vivo. ( A ) Blocking effect design of IL-4Rα blocking mAb in r Ts -serpin–treated or rmIL-4–treated peritoneal macrophages in vivo. Flow cytometry analysis of the F4/80 + ratio in nonlymphocyte subsets ( B ) and polarization in F4/80 + subsets ( C ) in rmIL-4–treated or r Ts -serpin–treated peritoneal cells coincubated with isotype control mAb or IL-4Rα blocking mAb. The values are the mean ± SD of eight independent experiments. Significant differences were analyzed by t test. ** p

    Techniques Used: Blocking Assay, In Vivo, Flow Cytometry

    3) Product Images from "The Anti-Inflammatory Immune Response in Early Trichinella spiralis Intestinal Infection Depends on Serine Protease Inhibitor–Mediated Alternative Activation of Macrophages"

    Article Title: The Anti-Inflammatory Immune Response in Early Trichinella spiralis Intestinal Infection Depends on Serine Protease Inhibitor–Mediated Alternative Activation of Macrophages

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.2000290

    The phenotypic effect of IL-4Rα blocking mAb on rmIL-4–treated or r Ts -serpin–treated peritoneal macrophages in vivo. ( A ) Blocking effect design of IL-4Rα blocking mAb in r Ts -serpin–treated or rmIL-4–treated peritoneal macrophages in vivo. Flow cytometry analysis of the F4/80 + ratio in nonlymphocyte subsets ( B ) and polarization in F4/80 + subsets ( C ) in rmIL-4–treated or r Ts -serpin–treated peritoneal cells coincubated with isotype control mAb or IL-4Rα blocking mAb. The values are the mean ± SD of eight independent experiments. Significant differences were analyzed by t test. ** p
    Figure Legend Snippet: The phenotypic effect of IL-4Rα blocking mAb on rmIL-4–treated or r Ts -serpin–treated peritoneal macrophages in vivo. ( A ) Blocking effect design of IL-4Rα blocking mAb in r Ts -serpin–treated or rmIL-4–treated peritoneal macrophages in vivo. Flow cytometry analysis of the F4/80 + ratio in nonlymphocyte subsets ( B ) and polarization in F4/80 + subsets ( C ) in rmIL-4–treated or r Ts -serpin–treated peritoneal cells coincubated with isotype control mAb or IL-4Rα blocking mAb. The values are the mean ± SD of eight independent experiments. Significant differences were analyzed by t test. ** p

    Techniques Used: Blocking Assay, In Vivo, Flow Cytometry

    4) Product Images from "Comparison of immature and mature bone marrow-derived dendritic cells by atomic force microscopy"

    Article Title: Comparison of immature and mature bone marrow-derived dendritic cells by atomic force microscopy

    Journal: Nanoscale Research Letters

    doi: 10.1186/1556-276X-6-455

    MHC-II expression and IL-12 production of immature and mature BMDCs . (A-D) Flow cytometry was used to detect CD11c and MHC-II molecule expression on the surface of the immature BMDCs treated with 10.0 μg/L of rmGM-CSF plus 10.0 μg/L of rmIL-4 as the control (A, C) or the mature BMDCs stimulated with 1.0 mg/L of LPS (B, D), which was displayed respectively by the scattered plots (A, B) and the single parameter diagrams (C, D). (E) The level of IL-12 secreted by the immature BMDCs or the mature BMDCs was measured by ELISA. * P
    Figure Legend Snippet: MHC-II expression and IL-12 production of immature and mature BMDCs . (A-D) Flow cytometry was used to detect CD11c and MHC-II molecule expression on the surface of the immature BMDCs treated with 10.0 μg/L of rmGM-CSF plus 10.0 μg/L of rmIL-4 as the control (A, C) or the mature BMDCs stimulated with 1.0 mg/L of LPS (B, D), which was displayed respectively by the scattered plots (A, B) and the single parameter diagrams (C, D). (E) The level of IL-12 secreted by the immature BMDCs or the mature BMDCs was measured by ELISA. * P

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    5) Product Images from "TET enzymes augment activation-induced deaminase (AID) expression via 5-hydroxymethylcytosine modifications at the Aicda superenhancer"

    Article Title: TET enzymes augment activation-induced deaminase (AID) expression via 5-hydroxymethylcytosine modifications at the Aicda superenhancer

    Journal: Science immunology

    doi: 10.1126/sciimmunol.aau7523

    TET proteins facilitate CSR. ( A ) Flowchart of in vivo experiment. f.p., footpad. ( B ) Top: Flow cytometric analysis of CD19 + GL7 + Fas + GC B cells at the draining popliteal lymph nodes from WT and Tet2/3 DKO mice after NP-OVA immunization as in (A). Bottom: Decreased IgG1-switched cells among GC B cells in Tet2/3 DKO (YFP + GC B–gated) compared with WT mice (GC B–gated). ( C to F ) Quantification of experiments shown in (B). Data shown are aggregated results from two independent experiments. Means and SEs are shown. WT, n = 11; DKO, n = 12. ( G ) Flowchart of in vitro IgG1 switching. Cells were labeled with Cell trace violet and activated for 4 days with LPS and IL-4. ( H and I ) Flow cytometry plots (H) and quantification (I) of IgG1-switched B cells in Tet2/3 DKO ( n = 4) and WT ( n = 4) mice. Representative of at least three independent experiments. ( J ) Circular γ1 transcript was quantified by qRT-PCR and normalized to Gapdh and then to the level of WT. Representative experiment of two independent experiments with three technical replicates. ( K ) Flowchart of in vitro IgA switching. Cells were activated for 5 days with anti-CD40, rmIL-4, rmIL-5, and rhTGFβ. ( L to N ) Flow cytometry plots (L) and quantification (M and N) of IgG1-switched (M) and IgA-switched cells (N). Representative of three independent experiments with three technical replicates. Statistical significance was calculated using unpaired two-tailed t test. * P
    Figure Legend Snippet: TET proteins facilitate CSR. ( A ) Flowchart of in vivo experiment. f.p., footpad. ( B ) Top: Flow cytometric analysis of CD19 + GL7 + Fas + GC B cells at the draining popliteal lymph nodes from WT and Tet2/3 DKO mice after NP-OVA immunization as in (A). Bottom: Decreased IgG1-switched cells among GC B cells in Tet2/3 DKO (YFP + GC B–gated) compared with WT mice (GC B–gated). ( C to F ) Quantification of experiments shown in (B). Data shown are aggregated results from two independent experiments. Means and SEs are shown. WT, n = 11; DKO, n = 12. ( G ) Flowchart of in vitro IgG1 switching. Cells were labeled with Cell trace violet and activated for 4 days with LPS and IL-4. ( H and I ) Flow cytometry plots (H) and quantification (I) of IgG1-switched B cells in Tet2/3 DKO ( n = 4) and WT ( n = 4) mice. Representative of at least three independent experiments. ( J ) Circular γ1 transcript was quantified by qRT-PCR and normalized to Gapdh and then to the level of WT. Representative experiment of two independent experiments with three technical replicates. ( K ) Flowchart of in vitro IgA switching. Cells were activated for 5 days with anti-CD40, rmIL-4, rmIL-5, and rhTGFβ. ( L to N ) Flow cytometry plots (L) and quantification (M and N) of IgG1-switched (M) and IgA-switched cells (N). Representative of three independent experiments with three technical replicates. Statistical significance was calculated using unpaired two-tailed t test. * P

    Techniques Used: In Vivo, Flow Cytometry, Mouse Assay, In Vitro, Labeling, Cytometry, Quantitative RT-PCR, Two Tailed Test

    6) Product Images from "Epithelial Cell-derived IL-25, but not Th17 cell-derived IL-17 or IL-17F, is Crucial for Murine Asthma"

    Article Title: Epithelial Cell-derived IL-25, but not Th17 cell-derived IL-17 or IL-17F, is Crucial for Murine Asthma

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1200461

    IL-25 can induce chemokine production by airway epithelial cells. The levels of CCL5 and CXCL1 in the culture supernatants and cell lysates of airway epithelial cells after stimulation with the indicated concentrations of rmIL-25, 100-ng/ml rmIL-4 or
    Figure Legend Snippet: IL-25 can induce chemokine production by airway epithelial cells. The levels of CCL5 and CXCL1 in the culture supernatants and cell lysates of airway epithelial cells after stimulation with the indicated concentrations of rmIL-25, 100-ng/ml rmIL-4 or

    Techniques Used:

    IL-25 can potently induce airway epithelial cell and eosinophil activation. A , B , Chemokine expression/production by MLE-12 cultured in the presence and absence of the indicated concentrations of rmIL-25, 100-ng/ml rmIL-4 or 100-ng/ml rmIL-13 for 4 hours
    Figure Legend Snippet: IL-25 can potently induce airway epithelial cell and eosinophil activation. A , B , Chemokine expression/production by MLE-12 cultured in the presence and absence of the indicated concentrations of rmIL-25, 100-ng/ml rmIL-4 or 100-ng/ml rmIL-13 for 4 hours

    Techniques Used: Activation Assay, Expressing, Cell Culture

    7) Product Images from "TLR9/MyD88 signaling is required for class switching to pathogenic IgG2a and 2b autoantibodies in SLE"

    Article Title: TLR9/MyD88 signaling is required for class switching to pathogenic IgG2a and 2b autoantibodies in SLE

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20052438

    Loss of IgG2a and 2b autoantibodies in MyD88-deficient mice was B cell intrinsic. Naive splenic B cells from the indicated genotypes were stimulated with 15 μg/ml CpG1826, 10 μg/ml anti-CD40, and 10 ng/ml rmIL-4 in vitro for 5 d and total Ig subclass concentrations in the supernatants were determined. Class switching to IgM was unchanged and to IgG1 increased in MyD88-deficient backgrounds. In contrast, switching to IgG2a, 2b, and 3 was significantly impaired.
    Figure Legend Snippet: Loss of IgG2a and 2b autoantibodies in MyD88-deficient mice was B cell intrinsic. Naive splenic B cells from the indicated genotypes were stimulated with 15 μg/ml CpG1826, 10 μg/ml anti-CD40, and 10 ng/ml rmIL-4 in vitro for 5 d and total Ig subclass concentrations in the supernatants were determined. Class switching to IgM was unchanged and to IgG1 increased in MyD88-deficient backgrounds. In contrast, switching to IgG2a, 2b, and 3 was significantly impaired.

    Techniques Used: Mouse Assay, In Vitro

    Loss of IgG2a and 2b autoantibodies in TLR9-deficient mice was B cell autonomous. (A) Naive splenic B cells from the indicated genotypes were stimulated with 15 μg/ml CpG1826, 10 μg/ml anti-CD40, and 10 ng/ml rmIL-4 in vitro for 5 d and total Ig subclass concentrations in the supernatants were determined. Class switching to IgM was unchanged and to IgG1 increased in TLR9-deficient backgrounds. In contrast, switching to IgG2a, 2b, and 3 was significantly impaired. (B) T-bet mRNA is not induced by CpG stimulation of B cells in the TLR9-deficient background. A portion of the cells in (A) were collected after 12 h of the indicated stimulation. mRNA and cDNA were prepared and used for real-time PCR with specific primers for T-bet and β-actin as an internal control. The ratio between TLR9-sufficient and -deficient B cells for T-bet mRNA was calculated after normalizing for β-actin. nd, not detected. Results from three independent experiments are shown as mean ± SD.
    Figure Legend Snippet: Loss of IgG2a and 2b autoantibodies in TLR9-deficient mice was B cell autonomous. (A) Naive splenic B cells from the indicated genotypes were stimulated with 15 μg/ml CpG1826, 10 μg/ml anti-CD40, and 10 ng/ml rmIL-4 in vitro for 5 d and total Ig subclass concentrations in the supernatants were determined. Class switching to IgM was unchanged and to IgG1 increased in TLR9-deficient backgrounds. In contrast, switching to IgG2a, 2b, and 3 was significantly impaired. (B) T-bet mRNA is not induced by CpG stimulation of B cells in the TLR9-deficient background. A portion of the cells in (A) were collected after 12 h of the indicated stimulation. mRNA and cDNA were prepared and used for real-time PCR with specific primers for T-bet and β-actin as an internal control. The ratio between TLR9-sufficient and -deficient B cells for T-bet mRNA was calculated after normalizing for β-actin. nd, not detected. Results from three independent experiments are shown as mean ± SD.

    Techniques Used: Mouse Assay, In Vitro, Real-time Polymerase Chain Reaction

    8) Product Images from "Hypoxia-induced mitogenic factor (FIZZ1/RELMα) induces endothelial cell apoptosis and subsequent interleukin-4-dependent pulmonary hypertension"

    Article Title: Hypoxia-induced mitogenic factor (FIZZ1/RELMα) induces endothelial cell apoptosis and subsequent interleukin-4-dependent pulmonary hypertension

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00279.2013

    Human resistin (RETN) and IL-4 induce EC apoptosis and activation, and the conditioned media from activated ECs enhances human pulmonary artery smooth muscle cell (HPSMC) proliferation. A : both RETN (200 ng/ml, 6 h) and IL-4 (50 ng/ml, 6 h) caused TUNEL-positive reactions in human PMVECs (×630 magnification). B : quantification of mean fluorescence intensity of RETN- and IL-4-induced TUNEL-positive cells is shown as a ratio of DAPI staining [ n = 8 low-magnification (×100) images from each group]. C : colocalization of RETN- or IL-4-induced Ang2 and vWF in PMVECs. Primary human PMVECs were treated with or without rhRETN (100 ng/ml) or rmIL-4 (50 ng/ml) for 24 h. Images are shown at ×630 magnification. D : quantification of mean fluorescence intensity of RETN- and IL-4-induced Ang2 expression is shown as a ratio of DAPI staining [ n = 8 low-magnification (×200) images from each group]. E : PMVECs were treated with 100 ng/ml rhRETN or 50 ng/ml rhIL-4 for 24 h, and the conditioned medium was added to serum- and growth factor-starved HPSMCs for 24 h. Serum-free smooth muscle basal medium (SmBM) was used as a negative control. rhRETN (100 ng/ml) or rhIL-4 (50 ng/ml) was added directly to HPSMCs for comparison. F : PMVECs were treated with 100 ng/ml rhRETN for 24 h, and ET-1 level in the conditioned medium was measured by ELISA. Values are means ± SE ( n = 4 separate culture cells/group, Student's t -test). Results are shown as means ± SE. **** P
    Figure Legend Snippet: Human resistin (RETN) and IL-4 induce EC apoptosis and activation, and the conditioned media from activated ECs enhances human pulmonary artery smooth muscle cell (HPSMC) proliferation. A : both RETN (200 ng/ml, 6 h) and IL-4 (50 ng/ml, 6 h) caused TUNEL-positive reactions in human PMVECs (×630 magnification). B : quantification of mean fluorescence intensity of RETN- and IL-4-induced TUNEL-positive cells is shown as a ratio of DAPI staining [ n = 8 low-magnification (×100) images from each group]. C : colocalization of RETN- or IL-4-induced Ang2 and vWF in PMVECs. Primary human PMVECs were treated with or without rhRETN (100 ng/ml) or rmIL-4 (50 ng/ml) for 24 h. Images are shown at ×630 magnification. D : quantification of mean fluorescence intensity of RETN- and IL-4-induced Ang2 expression is shown as a ratio of DAPI staining [ n = 8 low-magnification (×200) images from each group]. E : PMVECs were treated with 100 ng/ml rhRETN or 50 ng/ml rhIL-4 for 24 h, and the conditioned medium was added to serum- and growth factor-starved HPSMCs for 24 h. Serum-free smooth muscle basal medium (SmBM) was used as a negative control. rhRETN (100 ng/ml) or rhIL-4 (50 ng/ml) was added directly to HPSMCs for comparison. F : PMVECs were treated with 100 ng/ml rhRETN for 24 h, and ET-1 level in the conditioned medium was measured by ELISA. Values are means ± SE ( n = 4 separate culture cells/group, Student's t -test). Results are shown as means ± SE. **** P

    Techniques Used: Activation Assay, TUNEL Assay, Fluorescence, Staining, Expressing, Negative Control, Enzyme-linked Immunosorbent Assay

    HIMF and IL-4 activate VCAM-1 and angiopoietin 2 (Ang2) expression in mouse PMVECs. A–C : primary mouse PMVECs were treated with or without HIMF (20 nM) or rmIL-4 (50 ng/ml) for 24 h. A : VCAM-1 protein expression was analyzed in cell lysates ( left ). The ratio of VCAM-1 to β-actin is shown ( right ). *** P
    Figure Legend Snippet: HIMF and IL-4 activate VCAM-1 and angiopoietin 2 (Ang2) expression in mouse PMVECs. A–C : primary mouse PMVECs were treated with or without HIMF (20 nM) or rmIL-4 (50 ng/ml) for 24 h. A : VCAM-1 protein expression was analyzed in cell lysates ( left ). The ratio of VCAM-1 to β-actin is shown ( right ). *** P

    Techniques Used: Expressing

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    In Vitro:

    Article Title: T-bet regulates IgG class switching and pathogenic autoantibody production
    Article Snippet: Apoptosis was evaluated by exposing the cells for 24 h to 20 μg/ml soluble anti-mouse CD3 and anti-mouse CD28, 5 μg/ml dexamethasone (Sigma), or 1,200-J UV irradiation in a Stratalinker (Stratagene), followed by evaluation by the CaspACE Assay System (Promega). .. For in vitro analyses, purified mature B cells were isolated from spleen and lymph nodes by magnetic CD43 depletion (Miltenyi Biotec, Auburn, CA) and stimulated in RPMI/10% with 25 μg/ml LPS (Sigma) supplemented with recombinant murine IL-4 at 10 ng/ml, IFN-γ at 100 ng/ml, human TGF-β1 at 1 ng/ml (PeproTech, Rocky Hill, NJ), or murine IFN-α at 100 units/ml (R & D Systems). .. For retroviral infection studies, purified CD43-depleted mature B cells were stimulated by 25 μg/ml LPS for 24 h, followed by infection by a T-bet-GFP (green fluorescent protein) or control-GFP retrovirus ( ).

    Purification:

    Article Title: T-bet regulates IgG class switching and pathogenic autoantibody production
    Article Snippet: Apoptosis was evaluated by exposing the cells for 24 h to 20 μg/ml soluble anti-mouse CD3 and anti-mouse CD28, 5 μg/ml dexamethasone (Sigma), or 1,200-J UV irradiation in a Stratalinker (Stratagene), followed by evaluation by the CaspACE Assay System (Promega). .. For in vitro analyses, purified mature B cells were isolated from spleen and lymph nodes by magnetic CD43 depletion (Miltenyi Biotec, Auburn, CA) and stimulated in RPMI/10% with 25 μg/ml LPS (Sigma) supplemented with recombinant murine IL-4 at 10 ng/ml, IFN-γ at 100 ng/ml, human TGF-β1 at 1 ng/ml (PeproTech, Rocky Hill, NJ), or murine IFN-α at 100 units/ml (R & D Systems). .. For retroviral infection studies, purified CD43-depleted mature B cells were stimulated by 25 μg/ml LPS for 24 h, followed by infection by a T-bet-GFP (green fluorescent protein) or control-GFP retrovirus ( ).

    Isolation:

    Article Title: T-bet regulates IgG class switching and pathogenic autoantibody production
    Article Snippet: Apoptosis was evaluated by exposing the cells for 24 h to 20 μg/ml soluble anti-mouse CD3 and anti-mouse CD28, 5 μg/ml dexamethasone (Sigma), or 1,200-J UV irradiation in a Stratalinker (Stratagene), followed by evaluation by the CaspACE Assay System (Promega). .. For in vitro analyses, purified mature B cells were isolated from spleen and lymph nodes by magnetic CD43 depletion (Miltenyi Biotec, Auburn, CA) and stimulated in RPMI/10% with 25 μg/ml LPS (Sigma) supplemented with recombinant murine IL-4 at 10 ng/ml, IFN-γ at 100 ng/ml, human TGF-β1 at 1 ng/ml (PeproTech, Rocky Hill, NJ), or murine IFN-α at 100 units/ml (R & D Systems). .. For retroviral infection studies, purified CD43-depleted mature B cells were stimulated by 25 μg/ml LPS for 24 h, followed by infection by a T-bet-GFP (green fluorescent protein) or control-GFP retrovirus ( ).

    Derivative Assay:

    Article Title: Anti-inflammatory Microglia/Macrophages As a Potential Therapeutic Target in Brain Metastasis
    Article Snippet: Equal numbers of BV2 microglia or RAW 264.7 macrophages were seeded onto 12- or 96-well plates (Greiner Bio-one, CellStar). .. Cells were treated with 100 ng/ml LPS (E. coli derived, 0111:B4) (Sigma-Aldrich, UK) or 20 ng/ml murine recombinant IL-4 (Peprotech, UK) in DMEM for polarization to the pro- or anti-inflammatory phenotypes, respectively, or left untreated for the unpolarized control phenotype. .. At the end of the experiment, cells were fixed in 4% PFA (in PBS, pH 7.2) for immunofluorescence staining.

    Article Title: TLR7/8-agonist-loaded nanoparticles promote the polarization of tumour-associated macrophages to enhance cancer immunotherapy
    Article Snippet: The enrichment score for M1 cells was output back into the database and imported into KNIME to generate per-well and per-treatment averages. .. For transcriptional analysis, derived murine macrophages were treated with 10 ng mL−1 recombinant mouse IL-4 (PeproTech 214-14) for 24 hours to induce an M2-like polarization state and subsequently dosed with fresh media supplemented by pharmacologic drugs at the prescribed concentrations. ..

    Staining:

    Article Title: Epstein Barr Virus Interleukin 10 Suppresses Anti-inflammatory Phenotype in Human Monocytes
    Article Snippet: To differentiate monocytes into macrophages, cells were cultured with 50 ng/ml M-CSF (R & D systems). .. On day 6 cells were additionally stimulated with IFNγ (20 ng/ml, Peprotech), IL4 (20 ng/ml, Peprotech), hIL-10 (10 ng/ml), or vIL-10 (10 ng/ml) for 24 h. Surface markers were stained using following antibodies: CD14 Clone M5E2, CD163 Clone GHI/61, CD32 Clone FLI8.26 (BD Biosciences), CD16 Clone 3G8, CD64 Clone 10.1, HLA-DR Clone LN3, CD86 Clone IT2.2 (BioLegend). .. Cells were acquired on BD LSR II or BD Celesta (BD Biosciences) and data were analyzed using FlowJo (TreeStar, v10).

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  • 99
    PeproTech recombinant mouse il 4
    Rbpj -deficient alternatively activated macrophages are defective in suppressing T cell proliferation. (A) PECs from control, PBS-injected WT mice (filled gray histogram), and day-1 chitin-elicited PECs from WT mice (black line) and Rbpj cKO (grey line) were co-cultured with CFSE-labeled LN cells from OT-II transgenic mice and cognate OVA peptide. Proliferation of CD4 + cells was examined at 96 h of co-culture. Results are representative of three independent experiments. (B) CFSE-labeled OT-II transgenic LN cells were co-cultured with control (filled gray histogram) or <t>IL-4-treated</t> (10 ng/mL, black line) BMDMs from WT or Rbpj cKO mice and cognate OVA peptide. Proliferation of CD4 + cells was examined at 96 h of co-culture. Results are representative of two independent experiments
    Recombinant Mouse Il 4, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant mouse il 4/product/PeproTech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant mouse il 4 - by Bioz Stars, 2021-05
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    98
    PeproTech recombinant murine il 4
    Model of cell type-specific type regulation of IL-24 and IL-10 in NK cells and macrophages Pathways leading to IL-24 induction shown with red arrows. Pathways leading to IL-10 induction shown with green arrows. (A) ). We show that optimal LPS induction of IL-24 in macrophages is entirely independent of type I IFN receptor signaling but requires co-stimulation through the <t>IL-4/STAT6</t> pathway. LPS+IL-4 stimulation induces STAT6 recruitment to multiple regions within Il24 and STAT6-dependent chromatin remodeling of the Il24 locus. Enhanced stability of IL-24 mRNA in BMM occurs independently of the IL-4/STAT6 pathway (not shown). Of note, we also determined that type I IFN receptor-mediated regulation of IL-10 in BMM is independent of STAT4. (B) ).
    Recombinant Murine Il 4, supplied by PeproTech, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant murine il 4/product/PeproTech
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant murine il 4 - by Bioz Stars, 2021-05
    98/100 stars
      Buy from Supplier

    Image Search Results


    Rbpj -deficient alternatively activated macrophages are defective in suppressing T cell proliferation. (A) PECs from control, PBS-injected WT mice (filled gray histogram), and day-1 chitin-elicited PECs from WT mice (black line) and Rbpj cKO (grey line) were co-cultured with CFSE-labeled LN cells from OT-II transgenic mice and cognate OVA peptide. Proliferation of CD4 + cells was examined at 96 h of co-culture. Results are representative of three independent experiments. (B) CFSE-labeled OT-II transgenic LN cells were co-cultured with control (filled gray histogram) or IL-4-treated (10 ng/mL, black line) BMDMs from WT or Rbpj cKO mice and cognate OVA peptide. Proliferation of CD4 + cells was examined at 96 h of co-culture. Results are representative of two independent experiments

    Journal: Protein & Cell

    Article Title: RBP-J is required for M2 macrophage polarization in response to chitin and mediates expression of a subset of M2 genes

    doi: 10.1007/s13238-016-0248-7

    Figure Lengend Snippet: Rbpj -deficient alternatively activated macrophages are defective in suppressing T cell proliferation. (A) PECs from control, PBS-injected WT mice (filled gray histogram), and day-1 chitin-elicited PECs from WT mice (black line) and Rbpj cKO (grey line) were co-cultured with CFSE-labeled LN cells from OT-II transgenic mice and cognate OVA peptide. Proliferation of CD4 + cells was examined at 96 h of co-culture. Results are representative of three independent experiments. (B) CFSE-labeled OT-II transgenic LN cells were co-cultured with control (filled gray histogram) or IL-4-treated (10 ng/mL, black line) BMDMs from WT or Rbpj cKO mice and cognate OVA peptide. Proliferation of CD4 + cells was examined at 96 h of co-culture. Results are representative of two independent experiments

    Article Snippet: Recombinant mouse IL-4 was from Peprotech and used at 10 ng/mL unless otherwise noted.

    Techniques: Injection, Mouse Assay, Cell Culture, Labeling, Transgenic Assay, Co-Culture Assay

    A crucial role for Rbpj in regulating expression of a subset of M2 macrophage genes. BMDMs from WT and Rbpj cKO mice were stimulated with IL-4 (10 ng/mL) for the indicated times. (A) Immunoblot analysis of whole cell lysates using antibodies recognizing Arg1. Levels of SHP2 served as loading controls. Results are representative of three independent experiments. (B and C) mRNA expression (relative to GAPDH mRNA) was measured using quantitative real-time PCR. Cumulative results from three independent experiments are shown (errors bars indicate s.d.). P

    Journal: Protein & Cell

    Article Title: RBP-J is required for M2 macrophage polarization in response to chitin and mediates expression of a subset of M2 genes

    doi: 10.1007/s13238-016-0248-7

    Figure Lengend Snippet: A crucial role for Rbpj in regulating expression of a subset of M2 macrophage genes. BMDMs from WT and Rbpj cKO mice were stimulated with IL-4 (10 ng/mL) for the indicated times. (A) Immunoblot analysis of whole cell lysates using antibodies recognizing Arg1. Levels of SHP2 served as loading controls. Results are representative of three independent experiments. (B and C) mRNA expression (relative to GAPDH mRNA) was measured using quantitative real-time PCR. Cumulative results from three independent experiments are shown (errors bars indicate s.d.). P

    Article Snippet: Recombinant mouse IL-4 was from Peprotech and used at 10 ng/mL unless otherwise noted.

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    RBP-J regulates Arg1 expression in macrophages independently of STAT6 and IRF8. (A) BMDMs from WT and Rbpj cKO mice were stimulated with 100 ng/mL of IL-4 for the indicated times. Whole cell lysates were analyzed with immunoblotting using antibodies recognizing STAT6 phosphorylated on Tyr641 (pSTAT6). Levels of SHP2 served as loading controls. Results are representative of three independent experiments. (B) BMDMs from WT and Rbpj cKO mice were stimulated with 10 ng/mL of IL-4 and 1 ng/mL of LPS for the indicated times. Whole cell lysates were analyzed with immunoblotting using antibody recognizing IRF8. Levels of p38 served as loading controls. Results are representative of three independent experiments. (C) mRNA expression of Arg1 (relative to GAPDH mRNA) was measured in BMDMs from WT and Irf8 −/− mice. Cumulative results from two independent experiments (errors bars indicate s.d.) are shown. (D) BMDMs from WT and Irf8 −/− mice were stimulated with 10 ng/mL of IL-4 for the indicated times. Whole cell lysates were analyzed with immunoblotting using antibodies against Arg1 and IRF8. Levels of SHP2 served as loading controls. Results are representative of three independent experiments

    Journal: Protein & Cell

    Article Title: RBP-J is required for M2 macrophage polarization in response to chitin and mediates expression of a subset of M2 genes

    doi: 10.1007/s13238-016-0248-7

    Figure Lengend Snippet: RBP-J regulates Arg1 expression in macrophages independently of STAT6 and IRF8. (A) BMDMs from WT and Rbpj cKO mice were stimulated with 100 ng/mL of IL-4 for the indicated times. Whole cell lysates were analyzed with immunoblotting using antibodies recognizing STAT6 phosphorylated on Tyr641 (pSTAT6). Levels of SHP2 served as loading controls. Results are representative of three independent experiments. (B) BMDMs from WT and Rbpj cKO mice were stimulated with 10 ng/mL of IL-4 and 1 ng/mL of LPS for the indicated times. Whole cell lysates were analyzed with immunoblotting using antibody recognizing IRF8. Levels of p38 served as loading controls. Results are representative of three independent experiments. (C) mRNA expression of Arg1 (relative to GAPDH mRNA) was measured in BMDMs from WT and Irf8 −/− mice. Cumulative results from two independent experiments (errors bars indicate s.d.) are shown. (D) BMDMs from WT and Irf8 −/− mice were stimulated with 10 ng/mL of IL-4 for the indicated times. Whole cell lysates were analyzed with immunoblotting using antibodies against Arg1 and IRF8. Levels of SHP2 served as loading controls. Results are representative of three independent experiments

    Article Snippet: Recombinant mouse IL-4 was from Peprotech and used at 10 ng/mL unless otherwise noted.

    Techniques: Expressing, Mouse Assay

    Lipiodol transfected DC are immune modulatory. A . Day 7 bone marrow derived DC were cultured alone or transfected with mismatched siRNA, or IL-12p35-specific siRNA using 3 μl/culture lipiodol. Following a 24 hour incubation cells were plated at 1 × 10 6 in 6 well culture dishes and activated for 24 hours with 10 ng/ml LPS and 10 ng/ml TNF- α . Supernatant was harvested and analyzed by ELISA for IL-10 production. B . C57/BL6 DC were transfected with mismatched siRNA, IL-12p35-specific siRNA or lipiodol alone, irradiated (3,000 rad) and seeded in triplicate at various concentrations in a flat-bottom 96-well plate. Splenic T cells from BALB/c mice were added as responders (5 × 10 5 cells/well). The mixed lymphocytes were cultured for 72 h and proliferation was assessed by thymidine incorporation. C D . IFN-γ and IL-4 concentrations, respectively, were assessed from MLR cultures at 48 hours of incubation.

    Journal: Journal of Translational Medicine

    Article Title: A novel method of modifying immune responses by vaccination with lipiodol-siRNA mixtures

    doi: 10.1186/1479-5876-4-2

    Figure Lengend Snippet: Lipiodol transfected DC are immune modulatory. A . Day 7 bone marrow derived DC were cultured alone or transfected with mismatched siRNA, or IL-12p35-specific siRNA using 3 μl/culture lipiodol. Following a 24 hour incubation cells were plated at 1 × 10 6 in 6 well culture dishes and activated for 24 hours with 10 ng/ml LPS and 10 ng/ml TNF- α . Supernatant was harvested and analyzed by ELISA for IL-10 production. B . C57/BL6 DC were transfected with mismatched siRNA, IL-12p35-specific siRNA or lipiodol alone, irradiated (3,000 rad) and seeded in triplicate at various concentrations in a flat-bottom 96-well plate. Splenic T cells from BALB/c mice were added as responders (5 × 10 5 cells/well). The mixed lymphocytes were cultured for 72 h and proliferation was assessed by thymidine incorporation. C D . IFN-γ and IL-4 concentrations, respectively, were assessed from MLR cultures at 48 hours of incubation.

    Article Snippet: DC generation and siRNA transfection At Day 0, bone marrow cells were flushed from the femurs and tibias of C57/BL6 mice, washed and cultured in 6-well plates (Corning, NY) at 4 × 106 cells/well in 4 ml of complete medium (RPMI 1640 supplemented with 2 mM L-glutamine, 100 U/ml penicillin, 100 μg of streptomycin, 50 μM 2-ME, and 10% FCS (all from Life Technologies, Ontario, Canada) supplemented with recombinant GM-CSF (10 ng/ml; PeproTech, Rocky Hill, NJ) and recombinant mouse IL-4 (10 ng/ml; PeproTech).

    Techniques: Transfection, Derivative Assay, Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Irradiation, Mouse Assay

    In vivo immune modulation lipiodol/siRNA vaccine. C57/BL6 mice were immunized intradermally at the interior side of both hind legs with 100 μl of KLH or ovalbumin (1 μg/μl) emulsified in CFA in the presence or absence of 10 nMol siRNA and 10% lipiodol. After 14 days mice were sacrificed and T cell cytokine responses were assessed culturing purified CD4+ T cells with irradiated syngeneic splenocytes in triplicate and mixed with serial dilutions of antigen. IFN-γ in KLH (A) and ovalbumin (C) cultures, and IL-4 in KLH (B) and ovalbumin (D) cultures was assessed by ELISA.

    Journal: Journal of Translational Medicine

    Article Title: A novel method of modifying immune responses by vaccination with lipiodol-siRNA mixtures

    doi: 10.1186/1479-5876-4-2

    Figure Lengend Snippet: In vivo immune modulation lipiodol/siRNA vaccine. C57/BL6 mice were immunized intradermally at the interior side of both hind legs with 100 μl of KLH or ovalbumin (1 μg/μl) emulsified in CFA in the presence or absence of 10 nMol siRNA and 10% lipiodol. After 14 days mice were sacrificed and T cell cytokine responses were assessed culturing purified CD4+ T cells with irradiated syngeneic splenocytes in triplicate and mixed with serial dilutions of antigen. IFN-γ in KLH (A) and ovalbumin (C) cultures, and IL-4 in KLH (B) and ovalbumin (D) cultures was assessed by ELISA.

    Article Snippet: DC generation and siRNA transfection At Day 0, bone marrow cells were flushed from the femurs and tibias of C57/BL6 mice, washed and cultured in 6-well plates (Corning, NY) at 4 × 106 cells/well in 4 ml of complete medium (RPMI 1640 supplemented with 2 mM L-glutamine, 100 U/ml penicillin, 100 μg of streptomycin, 50 μM 2-ME, and 10% FCS (all from Life Technologies, Ontario, Canada) supplemented with recombinant GM-CSF (10 ng/ml; PeproTech, Rocky Hill, NJ) and recombinant mouse IL-4 (10 ng/ml; PeproTech).

    Techniques: In Vivo, Mouse Assay, Purification, Irradiation, Enzyme-linked Immunosorbent Assay

    IL-10 is mediates immune modulation by IL-12 silenced DC. A . MLR was performed with various concentrations of irradiated C57/BL6 stimulator DC transfected with siRNA to IL-12p35 or control transfected DC, and BALB/c responder T cells. 5 μl/ml of anti-IL-10 (JES5 2A5) antibody was added throughout the culture time. Supernatant was collected from 48-hour MLR cultures and assessed for IFN-γ or IL-4 ( B ) by ELISA.

    Journal: Journal of Translational Medicine

    Article Title: A novel method of modifying immune responses by vaccination with lipiodol-siRNA mixtures

    doi: 10.1186/1479-5876-4-2

    Figure Lengend Snippet: IL-10 is mediates immune modulation by IL-12 silenced DC. A . MLR was performed with various concentrations of irradiated C57/BL6 stimulator DC transfected with siRNA to IL-12p35 or control transfected DC, and BALB/c responder T cells. 5 μl/ml of anti-IL-10 (JES5 2A5) antibody was added throughout the culture time. Supernatant was collected from 48-hour MLR cultures and assessed for IFN-γ or IL-4 ( B ) by ELISA.

    Article Snippet: DC generation and siRNA transfection At Day 0, bone marrow cells were flushed from the femurs and tibias of C57/BL6 mice, washed and cultured in 6-well plates (Corning, NY) at 4 × 106 cells/well in 4 ml of complete medium (RPMI 1640 supplemented with 2 mM L-glutamine, 100 U/ml penicillin, 100 μg of streptomycin, 50 μM 2-ME, and 10% FCS (all from Life Technologies, Ontario, Canada) supplemented with recombinant GM-CSF (10 ng/ml; PeproTech, Rocky Hill, NJ) and recombinant mouse IL-4 (10 ng/ml; PeproTech).

    Techniques: Irradiation, Transfection, Enzyme-linked Immunosorbent Assay

    Model of cell type-specific type regulation of IL-24 and IL-10 in NK cells and macrophages Pathways leading to IL-24 induction shown with red arrows. Pathways leading to IL-10 induction shown with green arrows. (A) ). We show that optimal LPS induction of IL-24 in macrophages is entirely independent of type I IFN receptor signaling but requires co-stimulation through the IL-4/STAT6 pathway. LPS+IL-4 stimulation induces STAT6 recruitment to multiple regions within Il24 and STAT6-dependent chromatin remodeling of the Il24 locus. Enhanced stability of IL-24 mRNA in BMM occurs independently of the IL-4/STAT6 pathway (not shown). Of note, we also determined that type I IFN receptor-mediated regulation of IL-10 in BMM is independent of STAT4. (B) ).

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Cell-specific requirements for STAT proteins and type I IFN receptor signaling discretely regulate IL-24 and IL-10 expression in NK cells and macrophages

    doi: 10.4049/jimmunol.1701340

    Figure Lengend Snippet: Model of cell type-specific type regulation of IL-24 and IL-10 in NK cells and macrophages Pathways leading to IL-24 induction shown with red arrows. Pathways leading to IL-10 induction shown with green arrows. (A) ). We show that optimal LPS induction of IL-24 in macrophages is entirely independent of type I IFN receptor signaling but requires co-stimulation through the IL-4/STAT6 pathway. LPS+IL-4 stimulation induces STAT6 recruitment to multiple regions within Il24 and STAT6-dependent chromatin remodeling of the Il24 locus. Enhanced stability of IL-24 mRNA in BMM occurs independently of the IL-4/STAT6 pathway (not shown). Of note, we also determined that type I IFN receptor-mediated regulation of IL-10 in BMM is independent of STAT4. (B) ).

    Article Snippet: Recombinant murine IL-4, IL-12 (p70), and IL-13 were purchased from Peprotech (Rocky Hill, NJ, USA).

    Techniques:

    STAT6 is required for IL-4-induced IL-24 in NK cells and BMM NK cells (A, B) and BMM (C, D) were generated from WT (black) and Stat6 −/− mice (gray) in vitro and treated with the indicated stimuli for 6h (NK cells) or 3h (BMM). IL-24 (A, C) and IL-10 (B, D) mRNA expression was determined by RT-qPCR analysis. Data represent the mean ± SEM for 3 independent experiments with 3–5 mice per group, * p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Cell-specific requirements for STAT proteins and type I IFN receptor signaling discretely regulate IL-24 and IL-10 expression in NK cells and macrophages

    doi: 10.4049/jimmunol.1701340

    Figure Lengend Snippet: STAT6 is required for IL-4-induced IL-24 in NK cells and BMM NK cells (A, B) and BMM (C, D) were generated from WT (black) and Stat6 −/− mice (gray) in vitro and treated with the indicated stimuli for 6h (NK cells) or 3h (BMM). IL-24 (A, C) and IL-10 (B, D) mRNA expression was determined by RT-qPCR analysis. Data represent the mean ± SEM for 3 independent experiments with 3–5 mice per group, * p

    Article Snippet: Recombinant murine IL-4, IL-12 (p70), and IL-13 were purchased from Peprotech (Rocky Hill, NJ, USA).

    Techniques: Generated, Mouse Assay, In Vitro, Expressing, Quantitative RT-PCR

    IL-24 and IL-10 are co-expressed in cultured NK cells and BMM NK cells (A) were stimulated with the indicated cytokines for 6h and IL-24 (black) and IL-10 (gray) mRNA expression was determined by RT-qPCR analysis. Data represent the mean ± SEM of at least 4 independent experiments with 3–5 mice per group. (B) Kinetics of IL-24 and IL-10 mRNA expression in IL-2+IL-12-stimulated NK cells (one of two representative experiments with 3–5 mice per group). Data are presented as the relative fold increase (mRNA (Fold induction)) compared to non-stimulated cells which were assigned an arbitrary value of 1. (C, D) mRNA stability analysis of IL-24 (C) and IL-10 (D) in NK cells stimulated for 3h with IL-2 or IL-2+IL-12 (IL-2+12). Cells were then harvested (Control (0h)) or treated with actinomycin D (ActD) and incubated for and additional 2h (black bar) or 4h (gray bar). mRNA levels were measured by RT-qPCR and normalized to stimulated control cells that did not receive ActD (Control (0h). The data are presented as a percent of control of IL-24 or IL-10 mRNA expression which was assigned a value of 100%. Data are from one of two representative experiments with 3–5 mice per group. Dashed line indicates mRNA decay at 50% of the control. (E) Bone-marrow derived macrophages (BMM) were treated with the indicated stimuli for 3h and IL-24 and IL-10 mRNA expression was determined by RT-qPCR analysis. Data represent the mean ± SEM of at least 4 independent experiments with 3 mice per group. (F) Kinetics of IL-24 and IL-10 mRNA expression in LPS+IL-4-stimulated BMM (one of two representative experiments with 3 mice per group). Data are presented as the relative fold increase (mRNA (Fold induction)) compared to non-stimulated cells which were assigned an arbitrary value of 1. (G, H) mRNA stability analysis of IL-24 (G) and IL-10 (H) in BMMs stimulated for 2h with LPS or LPS+IL-4 (LPS+4). Cells were then harvested (Control (0h)) or treated with actinomycin D (ActD) and incubated for and additional 2h (black bar) or 4h (gray bar). mRNA levels were measured by RT-qPCR and normalized to stimulated control cells that did not receive ActD (Control (0h). The data are presented as a percent of control of IL-24 or IL-10 mRNA expression which was assigned a value of 100%. Data are from one of two representative experiments with 3 mice per group. Dashed line indicates mRNA decay at 50% of the control.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Cell-specific requirements for STAT proteins and type I IFN receptor signaling discretely regulate IL-24 and IL-10 expression in NK cells and macrophages

    doi: 10.4049/jimmunol.1701340

    Figure Lengend Snippet: IL-24 and IL-10 are co-expressed in cultured NK cells and BMM NK cells (A) were stimulated with the indicated cytokines for 6h and IL-24 (black) and IL-10 (gray) mRNA expression was determined by RT-qPCR analysis. Data represent the mean ± SEM of at least 4 independent experiments with 3–5 mice per group. (B) Kinetics of IL-24 and IL-10 mRNA expression in IL-2+IL-12-stimulated NK cells (one of two representative experiments with 3–5 mice per group). Data are presented as the relative fold increase (mRNA (Fold induction)) compared to non-stimulated cells which were assigned an arbitrary value of 1. (C, D) mRNA stability analysis of IL-24 (C) and IL-10 (D) in NK cells stimulated for 3h with IL-2 or IL-2+IL-12 (IL-2+12). Cells were then harvested (Control (0h)) or treated with actinomycin D (ActD) and incubated for and additional 2h (black bar) or 4h (gray bar). mRNA levels were measured by RT-qPCR and normalized to stimulated control cells that did not receive ActD (Control (0h). The data are presented as a percent of control of IL-24 or IL-10 mRNA expression which was assigned a value of 100%. Data are from one of two representative experiments with 3–5 mice per group. Dashed line indicates mRNA decay at 50% of the control. (E) Bone-marrow derived macrophages (BMM) were treated with the indicated stimuli for 3h and IL-24 and IL-10 mRNA expression was determined by RT-qPCR analysis. Data represent the mean ± SEM of at least 4 independent experiments with 3 mice per group. (F) Kinetics of IL-24 and IL-10 mRNA expression in LPS+IL-4-stimulated BMM (one of two representative experiments with 3 mice per group). Data are presented as the relative fold increase (mRNA (Fold induction)) compared to non-stimulated cells which were assigned an arbitrary value of 1. (G, H) mRNA stability analysis of IL-24 (G) and IL-10 (H) in BMMs stimulated for 2h with LPS or LPS+IL-4 (LPS+4). Cells were then harvested (Control (0h)) or treated with actinomycin D (ActD) and incubated for and additional 2h (black bar) or 4h (gray bar). mRNA levels were measured by RT-qPCR and normalized to stimulated control cells that did not receive ActD (Control (0h). The data are presented as a percent of control of IL-24 or IL-10 mRNA expression which was assigned a value of 100%. Data are from one of two representative experiments with 3 mice per group. Dashed line indicates mRNA decay at 50% of the control.

    Article Snippet: Recombinant murine IL-4, IL-12 (p70), and IL-13 were purchased from Peprotech (Rocky Hill, NJ, USA).

    Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Mouse Assay, Incubation, Derivative Assay

    Ig production in T-bet-deficient mice. ( A and B ) Serum Ig isotype titers were evaluated on 12-week-old Fas wt/wt ( A ) and Fas lpr/lpr ( B ) animals of the indicated T-bet genotypes by ELISA. ( C and D ) In vitro class switching was assayed in 48-h culture supernatants from purified CD43-depleted B cells. Three Fas wt/wt ( C ) and Fas lpr/lpr ( D ) animals of indicated T-bet genotypes were assayed for Ig isotype production by ELISA. Stimulatory conditions included: IgM, IgA, IgG2b, and IgG3, LPS 25 μg/ml; IgG1 and IgE, anti-CD40 2 μg/ml + 10 ng/ml recombinant mouse (rm) IL-4; IgG2a, LPS 25 μg/ml + 100 ng/ml rmIFN-γ. *, Undetectable by assay (less than 20 pg/ml).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: T-bet regulates IgG class switching and pathogenic autoantibody production

    doi: 10.1073/pnas.082114899

    Figure Lengend Snippet: Ig production in T-bet-deficient mice. ( A and B ) Serum Ig isotype titers were evaluated on 12-week-old Fas wt/wt ( A ) and Fas lpr/lpr ( B ) animals of the indicated T-bet genotypes by ELISA. ( C and D ) In vitro class switching was assayed in 48-h culture supernatants from purified CD43-depleted B cells. Three Fas wt/wt ( C ) and Fas lpr/lpr ( D ) animals of indicated T-bet genotypes were assayed for Ig isotype production by ELISA. Stimulatory conditions included: IgM, IgA, IgG2b, and IgG3, LPS 25 μg/ml; IgG1 and IgE, anti-CD40 2 μg/ml + 10 ng/ml recombinant mouse (rm) IL-4; IgG2a, LPS 25 μg/ml + 100 ng/ml rmIFN-γ. *, Undetectable by assay (less than 20 pg/ml).

    Article Snippet: For in vitro analyses, purified mature B cells were isolated from spleen and lymph nodes by magnetic CD43 depletion (Miltenyi Biotec, Auburn, CA) and stimulated in RPMI/10% with 25 μg/ml LPS (Sigma) supplemented with recombinant murine IL-4 at 10 ng/ml, IFN-γ at 100 ng/ml, human TGF-β1 at 1 ng/ml (PeproTech, Rocky Hill, NJ), or murine IFN-α at 100 units/ml (R & D Systems).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, In Vitro, Purification, Recombinant