il-6 Biolegend Search Results


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  • 99
    Thermo Fisher il 6 elisa kits
    Il 6 Elisa Kits, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 204 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 apc
    Il 6 Apc, supplied by BioLegend, used in various techniques. Bioz Stars score: 89/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend interleukin il 6
    Interleukin Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 93/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 antibodies
    AF-driven expansion of CD14 + HLA-DR -/low MDSC was dependent on <t>IL-6/IL10-STAT3</t> signal pathway PBMC from HD (n = 3) were treated with the AF (50% v/v) from OC patients (n=4) in the presence of neutralizing antibodies against IL-6 and/or IL-10 (A) or STAT3 inhibitor stattic (B) for 48 hours and then analyzed for the abundance of CD14 + HLA-DR -/low MDSC by flow cytometry. Addition of isotype antibody or DMSO was as controls. The representative dotplots were shown in left panel and the statistics were shown in right graph. The data are expressed as mean ± SEM of 4 biological replicates and representative of three independent experiments. *p
    Il 6 Antibodies, supplied by BioLegend, used in various techniques. Bioz Stars score: 78/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 elisa kits
    Effects of taraxasterol on the production of serum TNF- α and <t>IL-6</t> in mice with ethanol-induced liver injury. The mice were treated with taraxasterol (2.5, 5, and 10 mg/kg, respectively) or TPN and induced with EtOH as described in Materials and Methods. The levels of serum TNF- α and IL-6 were determined by <t>ELISA</t> kits. The values represent the means ± SEMs and are expressed as pg/ml of sera. ## P
    Il 6 Elisa Kits, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 elisa kit
    Effects of taraxasterol on the production of serum TNF- α and <t>IL-6</t> in mice with ethanol-induced liver injury. The mice were treated with taraxasterol (2.5, 5, and 10 mg/kg, respectively) or TPN and induced with EtOH as described in Materials and Methods. The levels of serum TNF- α and IL-6 were determined by <t>ELISA</t> kits. The values represent the means ± SEMs and are expressed as pg/ml of sera. ## P
    Il 6 Elisa Kit, supplied by BioLegend, used in various techniques. Bioz Stars score: 88/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend interleukin 6 il 6 levels
    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and <t>IL-6</t> ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P
    Interleukin 6 Il 6 Levels, supplied by BioLegend, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend apc anti mouse il 6
    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and <t>IL-6</t> ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P
    Apc Anti Mouse Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend pe anti mouse il 6
    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and <t>IL-6</t> ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P
    Pe Anti Mouse Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 95/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 specific elisa kit
    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and <t>IL-6</t> ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P
    Il 6 Specific Elisa Kit, supplied by BioLegend, used in various techniques. Bioz Stars score: 79/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend elisamax il 6 standard set
    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and <t>IL-6</t> ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P
    Elisamax Il 6 Standard Set, supplied by BioLegend, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend commercially available kits measuring il 6
    Human malignant ascites supernatant suppresses CD206 BiTE activity but not FRβ BiTE activity. a CFSE-stained MDMs were co-cultured with T cells (10:1 E:T ratio) and the indicated BiTEs, in the presence or absence of 50% ascitic supernatant from three patients (Patients 1, 2 and 5). 96 h later, percentage live MDMs was determined with propidium iodide staining and a Celigo image cytometer. b T cell activation was assessed by flow cytometric analysis of CD25 expression after 96 h of co-culture with MDMs and the indicated BiTEs, in the presence or absence of 50% ascitic supernatant from three patients (Patients 1, 2 and 5). c Quantities of <t>IL-6,</t> IL-10 and total (active and latent) TGF-β in malignant ascites fluid from six different patients, as determined by enzyme-linked immunosorbent assay. Normal serum (NS) pooled from three healthy donors was included as a control. d Quantities of soluble CD206 in malignant ascites fluid from nine different patients was determined by enzyme-linked immunosorbent assay. Pooled NS was used as a control. Each condition was measured in biological triplicate and represented as mean ± SD ( a - d ). Statistical significance was assessed by one-way ANOVA with Dunnett’s post-hoc analysis compared with “Pooled NS” ( c , d ), or two-way ANOVA followed by Bonferroni post-hoc analysis, with each treatment being compared to the “Mock” condition within the relevant group ( a and b ). (*, P
    Commercially Available Kits Measuring Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 70/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 elisa detection kit
    Pseudane-VII decreases LPS-induced pro-inflammatory cytokines. ( A ) BV-2 microglial cells were pretreated with diverse concentration (0.5, 1, 2.5, and 5 μM) of pseudane-VII for 2 h and then stimulated with LPS (200 ng/mL) for 6 h. Expression levels of mRNA were measured by RT-PCR. ( B ) Cells (2 × 10 4 cells/well) were seeded in 96-well culture plates and then were pretreated with pseudane-VII (0.5, 1, 2.5, and 5 μM) for 2 h. Pretreated BV-2 microglia cells were added with LPS (200 ng/mL) for 22 h. Supernatants were collected for cytokine release. IL-1β, TNF-α, and <t>IL-6</t> were measured by <t>ELISA</t> kit. Results are expressed as means (±SE) from three independent experiments. (** p
    Il 6 Elisa Detection Kit, supplied by BioLegend, used in various techniques. Bioz Stars score: 87/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend anti mouse il 6
    LRP1 deficient myeloid cells have a pro-inflammatory signature. Microglia (MG) and BMDM (MΦ) isolated from Cx3cr1 cre - Lrp1 fl/fl and Lrp1 fl/fl mice were treated with LPS (1 μg/mL) for 3 h (qPCR) or 24 h (ELISA). TNF-α expression in microglia was determined by qPCR ( a ) and ELISA ( b ). TNF-α expression in macrophages was determined by qPCR ( c ) and ELISA ( d ). Expression of IL-1β ( e ) and <t>IL-6</t> ( f ) in macrophages was determined by qPCR. g Transcript expression of IL-1β, IL-6, and TNF-α from the CNS of Cx3cr1 creER - Lrp1 fl/fl and Lrp1 fl/fl mice at the peak of EAE. 5, ** p
    Anti Mouse Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 92/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend anti murine il 6 antibody
    Levels of <t>IL-6</t> in lean and DIO WT mice
    Anti Murine Il 6 Antibody, supplied by BioLegend, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend mouse interleukin il 6
    Levels of <t>IL-6</t> in lean and DIO WT mice
    Mouse Interleukin Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 elisa max tm deluxe
    Levels of <t>IL-6</t> in lean and DIO WT mice
    Il 6 Elisa Max Tm Deluxe, supplied by BioLegend, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend purified anti mouse il 6
    Exogenous H 2 S alleviated inflammation response in ischemic myocardium from SOD1 KO mice. (a–b) Quantitative analysis of neutrophils (a) and macrophages (b) from SOD1 KO heart samples. (c) Immunohistochemistry of CD11b from SOD1 KO mice. Positive staining is shown in brown. Scale bar = 50 μ m. (d–e) Quantitative analysis of inflammatory cytokines <t>IL-6</t> (d) and TNF- α (e) in heart tissues. (f–g) Quantitative analysis of neutrophils (f) and monocytes (g) in blood samples from SOD1 KO mice. Values are mean ± SE, n = 6 in each group. ∗ p
    Purified Anti Mouse Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 80/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend biotin anti mouse il 6
    Exogenous H 2 S alleviated inflammation response in ischemic myocardium from SOD1 KO mice. (a–b) Quantitative analysis of neutrophils (a) and macrophages (b) from SOD1 KO heart samples. (c) Immunohistochemistry of CD11b from SOD1 KO mice. Positive staining is shown in brown. Scale bar = 50 μ m. (d–e) Quantitative analysis of inflammatory cytokines <t>IL-6</t> (d) and TNF- α (e) in heart tissues. (f–g) Quantitative analysis of neutrophils (f) and monocytes (g) in blood samples from SOD1 KO mice. Values are mean ± SE, n = 6 in each group. ∗ p
    Biotin Anti Mouse Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 6 elisa max standard kit
    The absence of secretory leukoprotease inhibitor (SLPI) in BM-derived eosinophils (BMEos) increases interleukin <t>(IL)-6</t> production and invasive activity. (A) I The production of IL-6 by B6 and Slpi −/− BMEos after lipopolysaccharide (LPS) stimulation for 12 h. (B) B6 and Slpi −/− BMEos were incubated with LPS (1 µg/ml) or IL-33 (0.1 µg/ml) for 12 h. BMEos were also incubated with TNP-OVA (1 ng/ml) for 12 h after the administration of 5 µg/ml anti-TNP-IgE. (A,B) IL-4, 6, and 13 levels in the supernatants of cells were measured by an <t>ELISA.</t> (C) The activities of tryptase and chymase prepared according to the methods described in Figure 2 B. (D) The amounts of eosinophil peroxidase (EPO) in B6 and Slpi −/− BMEos after the administration of the indicated stimulators. (E) Chemotactic assays of B6 and Slpi −/− eosinophils by LTB 4 (50 nM), CCL2 (50 nM), and CCL11 (10 nM). (F) Invasion assays using Matrigel in B6 and Slpi −/− eosinophils upon LPS (1 µg/ml) and CCL11 (10 nM) stimulation. All of the data are shown as the mean ± SEM of three different eosinophil cultures. * P
    Il 6 Elisa Max Standard Kit, supplied by BioLegend, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend mouse il 6
    C5a and C3a suppress IFN-β production in BMDCs WT BMDCs were pre-treated with either media, 50 nM C5a, or 500 nM C3a for 2 h, and then the cells were infected with Lm overnight. Cell-free supernatants were used to quantitate (A) IFN-β production or (B) <t>IL-6,</t> TNF-α, and MCP-1 production. (C) WT BMDCs were pre-treated with either media, 50 nM C5a, 100 nM C5aA, 500 nM C3a, or 100 nM C3aA for 2 h and then were incubated with 25 μg/ml c-di-AMP for 20 h. IFN-β was quantitated from cell-free supernatants. (D) C5aR−/− BMDCs were pre-treated with either media, 50 nM C5a, or 100 nM C5aA for 2 h and (E) C3aR−/− BMDCs were pre-treated with either media, 500 nM C3a, or 100 nM C3aA for 2 h, and then the cells were incubated with 25 μg/ml c-di-AMP for 20 h. IFN-β was quantitated from cell-free supernatants. All data are presented as mean pg/ml ± SEM. These data are pooled from three independent experiments. * P ≤ 0.017; ** P ≤ 0.009; *** P = 0.0006 by t test.
    Mouse Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 135 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend mouse recombinant il 6
    The expression of <t>IL-6</t> in cardiac mesenchymal fibroblasts is affected by aging. A ) Expression of IL-6 in quiescent fibroblasts derived from young and old MSCs were analyzed by quantitative PCR and by ELISA, 24 h after the cell cycle was synchronized. B ) Double immunofluorescence labeling of IL-6 (red) and DDR2 (green) in 3- and 30-mo-old mouse hearts. Double positive cells are visualized in yellow/orange color. Middle panel shows magnified image of the section marked by white squares. Bottom panel shows morphometric analysis of IL-6 + DDR2 + cells in heart sections. Nuclei were stained with DAPI. Scale bar, 20 μm. P
    Mouse Recombinant Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 87/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend human recombinant il 6
    JAK1 interacts Sirt1. A , JAK1 expression plasmid was co-transfected with or without FLAG-Sirt1 into HEK293 cells. The interaction of Sirt1 with JAK1 in the transfected cells was analyzed by co-immunoprecipitation with anti-FLAG antibody and by Western blotting with JAK1 antibody ( top panel ). The same membrane was reprobed with anti-Sirt1 ( middle panel ). JAK1 expression in the whole cell lysate was determined by Western blotting ( bottom panel ). B–D , MCF-10 ( B ), HCT116 ( C ), and MM.1s ( D ) cells were stimulated with or without 10 ng/ml <t>IL-6</t> and lysed with RIPA buffer. The interaction of Sirt1 and JAK1 was determined by immunoprecipitation of JAK1 using normal rabbit IgG ( rIgG ) as a control and by Western blotting with anti-Sirt1 antibody ( top panels ). The same membrane was stripped and reblotted with anti-JAK1 ( middle panels ). Sirt1 expression in the whole cell lysate was determined by Western blotting ( bottom panels ).
    Human Recombinant Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend antihuman il 6 capture
    JAK1 interacts Sirt1. A , JAK1 expression plasmid was co-transfected with or without FLAG-Sirt1 into HEK293 cells. The interaction of Sirt1 with JAK1 in the transfected cells was analyzed by co-immunoprecipitation with anti-FLAG antibody and by Western blotting with JAK1 antibody ( top panel ). The same membrane was reprobed with anti-Sirt1 ( middle panel ). JAK1 expression in the whole cell lysate was determined by Western blotting ( bottom panel ). B–D , MCF-10 ( B ), HCT116 ( C ), and MM.1s ( D ) cells were stimulated with or without 10 ng/ml <t>IL-6</t> and lysed with RIPA buffer. The interaction of Sirt1 and JAK1 was determined by immunoprecipitation of JAK1 using normal rabbit IgG ( rIgG ) as a control and by Western blotting with anti-Sirt1 antibody ( top panels ). The same membrane was stripped and reblotted with anti-JAK1 ( middle panels ). Sirt1 expression in the whole cell lysate was determined by Western blotting ( bottom panels ).
    Antihuman Il 6 Capture, supplied by BioLegend, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend leaf purified anti mouse il 6
    JAK1 interacts Sirt1. A , JAK1 expression plasmid was co-transfected with or without FLAG-Sirt1 into HEK293 cells. The interaction of Sirt1 with JAK1 in the transfected cells was analyzed by co-immunoprecipitation with anti-FLAG antibody and by Western blotting with JAK1 antibody ( top panel ). The same membrane was reprobed with anti-Sirt1 ( middle panel ). JAK1 expression in the whole cell lysate was determined by Western blotting ( bottom panel ). B–D , MCF-10 ( B ), HCT116 ( C ), and MM.1s ( D ) cells were stimulated with or without 10 ng/ml <t>IL-6</t> and lysed with RIPA buffer. The interaction of Sirt1 and JAK1 was determined by immunoprecipitation of JAK1 using normal rabbit IgG ( rIgG ) as a control and by Western blotting with anti-Sirt1 antibody ( top panels ). The same membrane was stripped and reblotted with anti-JAK1 ( middle panels ). Sirt1 expression in the whole cell lysate was determined by Western blotting ( bottom panels ).
    Leaf Purified Anti Mouse Il 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 89/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend rat anti mouse il 6 antibody
    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and <t>IL-6</t> ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P
    Rat Anti Mouse Il 6 Antibody, supplied by BioLegend, used in various techniques. Bioz Stars score: 82/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il6
    Pml regulates pro-inflammatory cytokines in MSCs. a ELISA array for the detection of soluble factors and cytokines in the supernatant of MSCs Pml +/+ or Pml −/− . The quantification of significantly different factors is shown in the lower chart. b Relative expression of cxcl1 (charts on the left) and <t>Il6</t> (charts on the right) in MSCs derived from Pml +/+ or Pml −/− mice (upper charts) or from Prrx1-Cre-Pml +/+ and Prrx1-Cre-Pml F /F mice (charts at the bottom). Cells of n = 3 mice each group have been pooled before RNA extraction. Data are shown as average ± SEM. c Western blot showing the reduction of Pml expression in HS5 cells treated with two independent shRNAs against Pml is shown on the left; the expression levels of cxcl1 and Il6 in cells silenced for the expression of Pml is shown in the charts on the right. Data are shown as average ± SEM
    Il6, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    BioLegend apc conjugated anti mouse il 6 antibody
    Pml regulates pro-inflammatory cytokines in MSCs. a ELISA array for the detection of soluble factors and cytokines in the supernatant of MSCs Pml +/+ or Pml −/− . The quantification of significantly different factors is shown in the lower chart. b Relative expression of cxcl1 (charts on the left) and <t>Il6</t> (charts on the right) in MSCs derived from Pml +/+ or Pml −/− mice (upper charts) or from Prrx1-Cre-Pml +/+ and Prrx1-Cre-Pml F /F mice (charts at the bottom). Cells of n = 3 mice each group have been pooled before RNA extraction. Data are shown as average ± SEM. c Western blot showing the reduction of Pml expression in HS5 cells treated with two independent shRNAs against Pml is shown on the left; the expression levels of cxcl1 and Il6 in cells silenced for the expression of Pml is shown in the charts on the right. Data are shown as average ± SEM
    Apc Conjugated Anti Mouse Il 6 Antibody, supplied by BioLegend, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/apc conjugated anti mouse il 6 antibody/product/BioLegend
    Average 79 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    apc conjugated anti mouse il 6 antibody - by Bioz Stars, 2020-01
    79/100 stars
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    81
    BioLegend phycoerythrin labeled anti mouse il 6 antibody
    iSN34 with CpG-B enhances <t>IL-6</t> production. Supernatants from stimulated cells were collected and IL-6 protein levels were measured by ELISA. Mouse splenocytes were harvested 48 h later and intracellular IL-6 protein levels were determined by ELISA. All assays were carried out at least three independent times in triplicate. Similar results were obtained from at least three different mice. Values are presented as mean + SD of three independent experiments, each performed in triplicate ( n = 9). Values with different letters (i.e., a, b, c, d, e, and f) were significantly different. **** p
    Phycoerythrin Labeled Anti Mouse Il 6 Antibody, supplied by BioLegend, used in various techniques. Bioz Stars score: 81/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phycoerythrin labeled anti mouse il 6 antibody/product/BioLegend
    Average 81 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    phycoerythrin labeled anti mouse il 6 antibody - by Bioz Stars, 2020-01
    81/100 stars
      Buy from Supplier

    Image Search Results


    AF-driven expansion of CD14 + HLA-DR -/low MDSC was dependent on IL-6/IL10-STAT3 signal pathway PBMC from HD (n = 3) were treated with the AF (50% v/v) from OC patients (n=4) in the presence of neutralizing antibodies against IL-6 and/or IL-10 (A) or STAT3 inhibitor stattic (B) for 48 hours and then analyzed for the abundance of CD14 + HLA-DR -/low MDSC by flow cytometry. Addition of isotype antibody or DMSO was as controls. The representative dotplots were shown in left panel and the statistics were shown in right graph. The data are expressed as mean ± SEM of 4 biological replicates and representative of three independent experiments. *p

    Journal: Oncotarget

    Article Title: Ascites-derived IL-6 and IL-10 synergistically expand CD14+HLA-DR-/low myeloid-derived suppressor cells in ovarian cancer patients

    doi: 10.18632/oncotarget.20164

    Figure Lengend Snippet: AF-driven expansion of CD14 + HLA-DR -/low MDSC was dependent on IL-6/IL10-STAT3 signal pathway PBMC from HD (n = 3) were treated with the AF (50% v/v) from OC patients (n=4) in the presence of neutralizing antibodies against IL-6 and/or IL-10 (A) or STAT3 inhibitor stattic (B) for 48 hours and then analyzed for the abundance of CD14 + HLA-DR -/low MDSC by flow cytometry. Addition of isotype antibody or DMSO was as controls. The representative dotplots were shown in left panel and the statistics were shown in right graph. The data are expressed as mean ± SEM of 4 biological replicates and representative of three independent experiments. *p

    Article Snippet: In some instances, PBMC were cultured with the AF in the presence of neutralizing antibodies against IL-6 and/or IL-10 (Biolegend) or STAT3 inhibitor stattic (Selleck) with isotype antibody or DMSO as controls.

    Techniques: Flow Cytometry, Cytometry

    The correlation between IL-6 and IL-10 levels and the abundance of CD14 + HLA-DR -/low MDSC in the ascites (A) IL-6 concentration in the sera and/or accompanying ascites from HD (n = 21) or OC (n = 11) patients. (B) IL-10 concentration in the sera and/or accompanying ascites from HD or OC patients. (C) The correlation between the abundance of CD14 + HLA-DR –/lo MDSC and IL-6 in ascites from OC patients (n = 11). (D) The correlation between the abundance of CD14 + HLA-DR –/lo MDSC and IL-10 in ascites from OC patients (n = 11). ***p

    Journal: Oncotarget

    Article Title: Ascites-derived IL-6 and IL-10 synergistically expand CD14+HLA-DR-/low myeloid-derived suppressor cells in ovarian cancer patients

    doi: 10.18632/oncotarget.20164

    Figure Lengend Snippet: The correlation between IL-6 and IL-10 levels and the abundance of CD14 + HLA-DR -/low MDSC in the ascites (A) IL-6 concentration in the sera and/or accompanying ascites from HD (n = 21) or OC (n = 11) patients. (B) IL-10 concentration in the sera and/or accompanying ascites from HD or OC patients. (C) The correlation between the abundance of CD14 + HLA-DR –/lo MDSC and IL-6 in ascites from OC patients (n = 11). (D) The correlation between the abundance of CD14 + HLA-DR –/lo MDSC and IL-10 in ascites from OC patients (n = 11). ***p

    Article Snippet: In some instances, PBMC were cultured with the AF in the presence of neutralizing antibodies against IL-6 and/or IL-10 (Biolegend) or STAT3 inhibitor stattic (Selleck) with isotype antibody or DMSO as controls.

    Techniques: Concentration Assay

    The correlation between relapse-free survival (RFS) and the abundance of CD14 + HLA-DR -/low MDSC and the levels of IL-6 and IL-10 in the OC patients (A) Kaplan-Meier plots showing the correlation between RFS and high or low levels (median as cutoff) of CD14 + HLA-DR –/lo MDSC in the PB (A) or accompanying ascites (B) , and IL-6 (C) or IL-10 (D) concentration in the accompanying ascites in the OC patients (n = 21 for PB and n = 11 for ascites) with p-Values determined by Mantel–Cox log-rank test.

    Journal: Oncotarget

    Article Title: Ascites-derived IL-6 and IL-10 synergistically expand CD14+HLA-DR-/low myeloid-derived suppressor cells in ovarian cancer patients

    doi: 10.18632/oncotarget.20164

    Figure Lengend Snippet: The correlation between relapse-free survival (RFS) and the abundance of CD14 + HLA-DR -/low MDSC and the levels of IL-6 and IL-10 in the OC patients (A) Kaplan-Meier plots showing the correlation between RFS and high or low levels (median as cutoff) of CD14 + HLA-DR –/lo MDSC in the PB (A) or accompanying ascites (B) , and IL-6 (C) or IL-10 (D) concentration in the accompanying ascites in the OC patients (n = 21 for PB and n = 11 for ascites) with p-Values determined by Mantel–Cox log-rank test.

    Article Snippet: In some instances, PBMC were cultured with the AF in the presence of neutralizing antibodies against IL-6 and/or IL-10 (Biolegend) or STAT3 inhibitor stattic (Selleck) with isotype antibody or DMSO as controls.

    Techniques: Concentration Assay

    Tumor-promoting effect of CC-MSCs on SW48 cells is reduced by the addition of anti-IL-6 antibody and the inhibitor of STAT3. a Transwell migration assay of SW48 cells exposed to CC-MSC-CM with or without anti-IL-6 antibody treatment (×100). b Colony-formation assay of SW48 cells treated with CC-MSC-CM in the presence or absence of anti-IL-6 antibody. c Wound-healing assay of SW48 cells exposed to CC-MSC-CM was performed in the presence or absence of anti-IL-6 antibody (magnification, ×50; scale bar: 500 μm). d Transwell migration assay of SW48 cells exposed to CC-MSC-CM with or without the STAT3 inhibitor (Stattic) treatment (×100). e Colony-formation assay of SW48 cells treated with CC-MSC-CM in the presence or absence of the STAT3 inhibitor (Stattic). f Wound-healing assay of SW48 cells exposed to CC-MSC-CM was performed in the presence or absence of the STAT3 inhibitor (Stattic) (magnification, 50X; scale bar: 500 μm). * P

    Journal: Cell Death & Disease

    Article Title: Human colorectal cancer-derived mesenchymal stem cells promote colorectal cancer progression through IL-6/JAK2/STAT3 signaling

    doi: 10.1038/s41419-017-0176-3

    Figure Lengend Snippet: Tumor-promoting effect of CC-MSCs on SW48 cells is reduced by the addition of anti-IL-6 antibody and the inhibitor of STAT3. a Transwell migration assay of SW48 cells exposed to CC-MSC-CM with or without anti-IL-6 antibody treatment (×100). b Colony-formation assay of SW48 cells treated with CC-MSC-CM in the presence or absence of anti-IL-6 antibody. c Wound-healing assay of SW48 cells exposed to CC-MSC-CM was performed in the presence or absence of anti-IL-6 antibody (magnification, ×50; scale bar: 500 μm). d Transwell migration assay of SW48 cells exposed to CC-MSC-CM with or without the STAT3 inhibitor (Stattic) treatment (×100). e Colony-formation assay of SW48 cells treated with CC-MSC-CM in the presence or absence of the STAT3 inhibitor (Stattic). f Wound-healing assay of SW48 cells exposed to CC-MSC-CM was performed in the presence or absence of the STAT3 inhibitor (Stattic) (magnification, 50X; scale bar: 500 μm). * P

    Article Snippet: Stattic (MedChem Express, NJ, USA), a specific chemical inhibitor of STAT3; Anti-IL-6 antibody (Biolegend, CA, USA), a neutralizing antibody for IL-6.

    Techniques: Transwell Migration Assay, Colony Assay, Wound Healing Assay

    IL-6 secreted by CC-MSCs enhances the proliferation, migration and invasion of colorectal cancer cells through IL-6/JAK2/STAT3 signaling. a The levels of various factors in the cell-free culture supernatants were measured using Bio-Plex cytokine arrays. b Quantitative analysis of IL-6 levels using enzyme-linked immunosorbent assay (ELISA). The conditioned media from the cultured CC-MSCs and SW48 cells were collected to detect the levels of IL-6, and representative results from one of the three independent experiments are presented. c Transwell migration (top) and invasion (bottom) assay of SW48 cells with or without 10 ng/mL recombinant IL-6 treatment (×100). d Colony-formation assay of SW48 cells treated with or without 10 ng/mL recombinant IL-6. e Wound-healing assay of SW48 cells in the presence or absence of recombinant IL-6 (magnification, ×50; scale bar: 500 μm). *** P

    Journal: Cell Death & Disease

    Article Title: Human colorectal cancer-derived mesenchymal stem cells promote colorectal cancer progression through IL-6/JAK2/STAT3 signaling

    doi: 10.1038/s41419-017-0176-3

    Figure Lengend Snippet: IL-6 secreted by CC-MSCs enhances the proliferation, migration and invasion of colorectal cancer cells through IL-6/JAK2/STAT3 signaling. a The levels of various factors in the cell-free culture supernatants were measured using Bio-Plex cytokine arrays. b Quantitative analysis of IL-6 levels using enzyme-linked immunosorbent assay (ELISA). The conditioned media from the cultured CC-MSCs and SW48 cells were collected to detect the levels of IL-6, and representative results from one of the three independent experiments are presented. c Transwell migration (top) and invasion (bottom) assay of SW48 cells with or without 10 ng/mL recombinant IL-6 treatment (×100). d Colony-formation assay of SW48 cells treated with or without 10 ng/mL recombinant IL-6. e Wound-healing assay of SW48 cells in the presence or absence of recombinant IL-6 (magnification, ×50; scale bar: 500 μm). *** P

    Article Snippet: Stattic (MedChem Express, NJ, USA), a specific chemical inhibitor of STAT3; Anti-IL-6 antibody (Biolegend, CA, USA), a neutralizing antibody for IL-6.

    Techniques: Migration, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant, Colony Assay, Wound Healing Assay

    Effects of taraxasterol on the production of serum TNF- α and IL-6 in mice with ethanol-induced liver injury. The mice were treated with taraxasterol (2.5, 5, and 10 mg/kg, respectively) or TPN and induced with EtOH as described in Materials and Methods. The levels of serum TNF- α and IL-6 were determined by ELISA kits. The values represent the means ± SEMs and are expressed as pg/ml of sera. ## P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Protective Effects of Taraxasterol against Ethanol-Induced Liver Injury by Regulating CYP2E1/Nrf2/HO-1 and NF-κB Signaling Pathways in Mice

    doi: 10.1155/2018/8284107

    Figure Lengend Snippet: Effects of taraxasterol on the production of serum TNF- α and IL-6 in mice with ethanol-induced liver injury. The mice were treated with taraxasterol (2.5, 5, and 10 mg/kg, respectively) or TPN and induced with EtOH as described in Materials and Methods. The levels of serum TNF- α and IL-6 were determined by ELISA kits. The values represent the means ± SEMs and are expressed as pg/ml of sera. ## P

    Article Snippet: Mouse TNF-α and IL-6 ELISA kits were purchased from BioLegend Inc. (San Diego, CA, USA).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Effect of IFN-τ on IL-10 and IL-6 production in EECs ELISA was performed to detect (A) IL-10 and (B) IL-6 expression in EECs treated with control DMSO (CG) or IFN-τ. * P

    Journal: Oncotarget

    Article Title: Interferon-τ increases BoLA-I for implantation during early pregnancy in dairy cows

    doi: 10.18632/oncotarget.19282

    Figure Lengend Snippet: Effect of IFN-τ on IL-10 and IL-6 production in EECs ELISA was performed to detect (A) IL-10 and (B) IL-6 expression in EECs treated with control DMSO (CG) or IFN-τ. * P

    Article Snippet: Bovine IL-10 and IL-6 enzyme-linked immunosorbent assay (ELISA) kits were obtained from Biolegend (San Diego, CA, USA).

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing

    Production of IL-10 and IL-6 in endometrial tissues IL-10 (A) and IL-6 (B) expression in endometrial tissues of pregnant and non-pregnant cows was measured by ELISA. Columns represent the means. Bars represent the S.E. * P

    Journal: Oncotarget

    Article Title: Interferon-τ increases BoLA-I for implantation during early pregnancy in dairy cows

    doi: 10.18632/oncotarget.19282

    Figure Lengend Snippet: Production of IL-10 and IL-6 in endometrial tissues IL-10 (A) and IL-6 (B) expression in endometrial tissues of pregnant and non-pregnant cows was measured by ELISA. Columns represent the means. Bars represent the S.E. * P

    Article Snippet: Bovine IL-10 and IL-6 enzyme-linked immunosorbent assay (ELISA) kits were obtained from Biolegend (San Diego, CA, USA).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Role of TNF secretion in LegC4-mediated attenuation of L. pneumophila replication. (A and B) ELISA quantification of TNF (A) or IL-6 (B) secretion from wild-type BMDMs infected with L. pneumophila Δ flaA Δ legC4 (pEV) and Δ flaA Δ

    Journal: Journal of Bacteriology

    Article Title: Potentiation of Cytokine-Mediated Restriction of Legionella Intracellular Replication by a Dot/Icm-Translocated Effector

    doi: 10.1128/JB.00755-18

    Figure Lengend Snippet: Role of TNF secretion in LegC4-mediated attenuation of L. pneumophila replication. (A and B) ELISA quantification of TNF (A) or IL-6 (B) secretion from wild-type BMDMs infected with L. pneumophila Δ flaA Δ legC4 (pEV) and Δ flaA Δ

    Article Snippet: Supernatants were either used fresh or stored at −20°C for up to 1 week, followed by quantification of TNF or IL-6 using a mouse TNF or IL-6 enzyme-linked immunosorbent assay (ELISA) kit (BioLegend) following the manufacturer’s instructions.

    Techniques: Enzyme-linked Immunosorbent Assay, Infection

    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and IL-6 ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P

    Journal: Scientific Reports

    Article Title: Thioacetamide-induced liver damage and thrombocytopenia is associated with induction of antiplatelet autoantibody in mice

    doi: 10.1038/s41598-019-53977-7

    Figure Lengend Snippet: B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and IL-6 ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P

    Article Snippet: The ELISA kit used for analysing plasma levels of proinflammatory cytokine tumour necrosis factor-α (TNF-α), high mobility group protein B1 (HMGB-1), and interleukin-6 (IL-6) levels was purchased from BioLegend (San Diego, CA, USA).

    Techniques: Mouse Assay, Expressing, AST Assay

    Human malignant ascites supernatant suppresses CD206 BiTE activity but not FRβ BiTE activity. a CFSE-stained MDMs were co-cultured with T cells (10:1 E:T ratio) and the indicated BiTEs, in the presence or absence of 50% ascitic supernatant from three patients (Patients 1, 2 and 5). 96 h later, percentage live MDMs was determined with propidium iodide staining and a Celigo image cytometer. b T cell activation was assessed by flow cytometric analysis of CD25 expression after 96 h of co-culture with MDMs and the indicated BiTEs, in the presence or absence of 50% ascitic supernatant from three patients (Patients 1, 2 and 5). c Quantities of IL-6, IL-10 and total (active and latent) TGF-β in malignant ascites fluid from six different patients, as determined by enzyme-linked immunosorbent assay. Normal serum (NS) pooled from three healthy donors was included as a control. d Quantities of soluble CD206 in malignant ascites fluid from nine different patients was determined by enzyme-linked immunosorbent assay. Pooled NS was used as a control. Each condition was measured in biological triplicate and represented as mean ± SD ( a - d ). Statistical significance was assessed by one-way ANOVA with Dunnett’s post-hoc analysis compared with “Pooled NS” ( c , d ), or two-way ANOVA followed by Bonferroni post-hoc analysis, with each treatment being compared to the “Mock” condition within the relevant group ( a and b ). (*, P

    Journal: Journal for Immunotherapy of Cancer

    Article Title: Bi- and tri-valent T cell engagers deplete tumour-associated macrophages in cancer patient samples

    doi: 10.1186/s40425-019-0807-6

    Figure Lengend Snippet: Human malignant ascites supernatant suppresses CD206 BiTE activity but not FRβ BiTE activity. a CFSE-stained MDMs were co-cultured with T cells (10:1 E:T ratio) and the indicated BiTEs, in the presence or absence of 50% ascitic supernatant from three patients (Patients 1, 2 and 5). 96 h later, percentage live MDMs was determined with propidium iodide staining and a Celigo image cytometer. b T cell activation was assessed by flow cytometric analysis of CD25 expression after 96 h of co-culture with MDMs and the indicated BiTEs, in the presence or absence of 50% ascitic supernatant from three patients (Patients 1, 2 and 5). c Quantities of IL-6, IL-10 and total (active and latent) TGF-β in malignant ascites fluid from six different patients, as determined by enzyme-linked immunosorbent assay. Normal serum (NS) pooled from three healthy donors was included as a control. d Quantities of soluble CD206 in malignant ascites fluid from nine different patients was determined by enzyme-linked immunosorbent assay. Pooled NS was used as a control. Each condition was measured in biological triplicate and represented as mean ± SD ( a - d ). Statistical significance was assessed by one-way ANOVA with Dunnett’s post-hoc analysis compared with “Pooled NS” ( c , d ), or two-way ANOVA followed by Bonferroni post-hoc analysis, with each treatment being compared to the “Mock” condition within the relevant group ( a and b ). (*, P

    Article Snippet: Enzyme-linked immunosorbent assay (ELISA) ELISAs were performed with commercially-available kits measuring IL-6 (Biolegend, UK, #30504), IL-10 (Biolegend, UK, #430604), IFN-γ (Biolegend, UK, #430104), TGF beta-1 Human/Mouse ELISA Kit (Invitrogen, UK, #88–8350-88) and CD206 (RayBiotech, USA, #ELH-MMR-1), following manufacturers’ instructions.

    Techniques: Activity Assay, Staining, Cell Culture, Cytometry, Activation Assay, Flow Cytometry, Expressing, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

    Pseudane-VII decreases LPS-induced pro-inflammatory cytokines. ( A ) BV-2 microglial cells were pretreated with diverse concentration (0.5, 1, 2.5, and 5 μM) of pseudane-VII for 2 h and then stimulated with LPS (200 ng/mL) for 6 h. Expression levels of mRNA were measured by RT-PCR. ( B ) Cells (2 × 10 4 cells/well) were seeded in 96-well culture plates and then were pretreated with pseudane-VII (0.5, 1, 2.5, and 5 μM) for 2 h. Pretreated BV-2 microglia cells were added with LPS (200 ng/mL) for 22 h. Supernatants were collected for cytokine release. IL-1β, TNF-α, and IL-6 were measured by ELISA kit. Results are expressed as means (±SE) from three independent experiments. (** p

    Journal: Molecules

    Article Title: Pseudane-VII Regulates LPS-Induced Neuroinflammation in Brain Microglia Cells through the Inhibition of iNOS Expression

    doi: 10.3390/molecules23123196

    Figure Lengend Snippet: Pseudane-VII decreases LPS-induced pro-inflammatory cytokines. ( A ) BV-2 microglial cells were pretreated with diverse concentration (0.5, 1, 2.5, and 5 μM) of pseudane-VII for 2 h and then stimulated with LPS (200 ng/mL) for 6 h. Expression levels of mRNA were measured by RT-PCR. ( B ) Cells (2 × 10 4 cells/well) were seeded in 96-well culture plates and then were pretreated with pseudane-VII (0.5, 1, 2.5, and 5 μM) for 2 h. Pretreated BV-2 microglia cells were added with LPS (200 ng/mL) for 22 h. Supernatants were collected for cytokine release. IL-1β, TNF-α, and IL-6 were measured by ELISA kit. Results are expressed as means (±SE) from three independent experiments. (** p

    Article Snippet: TNF-α, IL-1β, and IL-6 ELISA detection kit were purchased from BioLegend (San Diego, CA, USA).

    Techniques: Concentration Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    LRP1 deficient myeloid cells have a pro-inflammatory signature. Microglia (MG) and BMDM (MΦ) isolated from Cx3cr1 cre - Lrp1 fl/fl and Lrp1 fl/fl mice were treated with LPS (1 μg/mL) for 3 h (qPCR) or 24 h (ELISA). TNF-α expression in microglia was determined by qPCR ( a ) and ELISA ( b ). TNF-α expression in macrophages was determined by qPCR ( c ) and ELISA ( d ). Expression of IL-1β ( e ) and IL-6 ( f ) in macrophages was determined by qPCR. g Transcript expression of IL-1β, IL-6, and TNF-α from the CNS of Cx3cr1 creER - Lrp1 fl/fl and Lrp1 fl/fl mice at the peak of EAE. 5, ** p

    Journal: Acta Neuropathologica Communications

    Article Title: LRP1 expression in microglia is protective during CNS autoimmunity

    doi: 10.1186/s40478-016-0343-2

    Figure Lengend Snippet: LRP1 deficient myeloid cells have a pro-inflammatory signature. Microglia (MG) and BMDM (MΦ) isolated from Cx3cr1 cre - Lrp1 fl/fl and Lrp1 fl/fl mice were treated with LPS (1 μg/mL) for 3 h (qPCR) or 24 h (ELISA). TNF-α expression in microglia was determined by qPCR ( a ) and ELISA ( b ). TNF-α expression in macrophages was determined by qPCR ( c ) and ELISA ( d ). Expression of IL-1β ( e ) and IL-6 ( f ) in macrophages was determined by qPCR. g Transcript expression of IL-1β, IL-6, and TNF-α from the CNS of Cx3cr1 creER - Lrp1 fl/fl and Lrp1 fl/fl mice at the peak of EAE. 5, ** p

    Article Snippet: Antibodies used were: anti-mouse IL-6 (MP5-20 F3, Biolegend) 0.5 μg/mL; biotin anti-mouse IL-6 (MP5-32C11, Biolegend) 1 μg/mL; anti-mouse TNF-α (R & D systems, AF-410-NA) 0.5 μg/mL; biotin anti-mouse TNF-α (R & D systems, BAF410) 0.25 μg/mL.

    Techniques: Isolation, Mouse Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing

    LRP1 is an inhibitor of NF-kB through the MyD88 pathway. a Macrophages isolated from Cx3cr1 cre - Lrp1 fl/fl (LRP1-) and Lrp1 fl/fl (LRP1+) mice were treated with LPS (1 μg/ml) and the expression of p-p65, p65, LRP1 and actin were determined by immunoblot. Representative of 3 independent experiments). b LRP1+ and LRP1- macrophages were treated with TLR ligands and expression of IL-6 was determined by qPCR analysis. * p

    Journal: Acta Neuropathologica Communications

    Article Title: LRP1 expression in microglia is protective during CNS autoimmunity

    doi: 10.1186/s40478-016-0343-2

    Figure Lengend Snippet: LRP1 is an inhibitor of NF-kB through the MyD88 pathway. a Macrophages isolated from Cx3cr1 cre - Lrp1 fl/fl (LRP1-) and Lrp1 fl/fl (LRP1+) mice were treated with LPS (1 μg/ml) and the expression of p-p65, p65, LRP1 and actin were determined by immunoblot. Representative of 3 independent experiments). b LRP1+ and LRP1- macrophages were treated with TLR ligands and expression of IL-6 was determined by qPCR analysis. * p

    Article Snippet: Antibodies used were: anti-mouse IL-6 (MP5-20 F3, Biolegend) 0.5 μg/mL; biotin anti-mouse IL-6 (MP5-32C11, Biolegend) 1 μg/mL; anti-mouse TNF-α (R & D systems, AF-410-NA) 0.5 μg/mL; biotin anti-mouse TNF-α (R & D systems, BAF410) 0.25 μg/mL.

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

    Levels of IL-6 in lean and DIO WT mice

    Journal: Cytokine

    Article Title: Hematological and acute-phase responses to diet-induced obesity in IL-6 KO mice

    doi: 10.1016/j.cyto.2011.09.015

    Figure Lengend Snippet: Levels of IL-6 in lean and DIO WT mice

    Article Snippet: For IL-6 neutralization experiments, lean and DIO WT mice were injected ip with 1 mg of an anti-murine IL-6 antibody (Biolegend, San Diego CA) previously showed to effectively neutralize IL-6 activity in vivo [ , ].

    Techniques: Mouse Assay

    Exogenous H 2 S alleviated inflammation response in ischemic myocardium from SOD1 KO mice. (a–b) Quantitative analysis of neutrophils (a) and macrophages (b) from SOD1 KO heart samples. (c) Immunohistochemistry of CD11b from SOD1 KO mice. Positive staining is shown in brown. Scale bar = 50 μ m. (d–e) Quantitative analysis of inflammatory cytokines IL-6 (d) and TNF- α (e) in heart tissues. (f–g) Quantitative analysis of neutrophils (f) and monocytes (g) in blood samples from SOD1 KO mice. Values are mean ± SE, n = 6 in each group. ∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hydrogen Sulfide Alleviates Acute Myocardial Ischemia Injury by Modulating Autophagy and Inflammation Response under Oxidative Stress

    doi: 10.1155/2018/3402809

    Figure Lengend Snippet: Exogenous H 2 S alleviated inflammation response in ischemic myocardium from SOD1 KO mice. (a–b) Quantitative analysis of neutrophils (a) and macrophages (b) from SOD1 KO heart samples. (c) Immunohistochemistry of CD11b from SOD1 KO mice. Positive staining is shown in brown. Scale bar = 50 μ m. (d–e) Quantitative analysis of inflammatory cytokines IL-6 (d) and TNF- α (e) in heart tissues. (f–g) Quantitative analysis of neutrophils (f) and monocytes (g) in blood samples from SOD1 KO mice. Values are mean ± SE, n = 6 in each group. ∗ p

    Article Snippet: IL-6 was detected using a rat anti-mouse IL-6 monoclonal antibody (504502, BioLegend, San Diego, CA, USA) for capture and biotinylated rat anti-mouse IL-6 monoclonal antibody (504602, BioLegend, San Diego, CA, USA) for detection.

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    SOD1 KO mice subjected to AMI showed severe inflammation response in the ischemic myocardium. (a) Representative FACS results of inflammatory cell counting from heart samples. (b–c) Quantitative analysis of neutrophils (b) and macrophages (c) in heart tissues. (d) Immunohistochemistry of CD11b (marker of both neutrophils and macrophages). Positive staining is shown in brown. Scale bar = 50 μ m. (e–f) Quantitative analysis of inflammatory cytokines IL-6 (e) and TNF- α (f) in heart tissues. (g–h) Quantitative analysis of neutrophils (g) and monocytes (h) in blood samples. Values are mean ± SE, n = 6 in each group. ∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hydrogen Sulfide Alleviates Acute Myocardial Ischemia Injury by Modulating Autophagy and Inflammation Response under Oxidative Stress

    doi: 10.1155/2018/3402809

    Figure Lengend Snippet: SOD1 KO mice subjected to AMI showed severe inflammation response in the ischemic myocardium. (a) Representative FACS results of inflammatory cell counting from heart samples. (b–c) Quantitative analysis of neutrophils (b) and macrophages (c) in heart tissues. (d) Immunohistochemistry of CD11b (marker of both neutrophils and macrophages). Positive staining is shown in brown. Scale bar = 50 μ m. (e–f) Quantitative analysis of inflammatory cytokines IL-6 (e) and TNF- α (f) in heart tissues. (g–h) Quantitative analysis of neutrophils (g) and monocytes (h) in blood samples. Values are mean ± SE, n = 6 in each group. ∗ p

    Article Snippet: IL-6 was detected using a rat anti-mouse IL-6 monoclonal antibody (504502, BioLegend, San Diego, CA, USA) for capture and biotinylated rat anti-mouse IL-6 monoclonal antibody (504602, BioLegend, San Diego, CA, USA) for detection.

    Techniques: Mouse Assay, FACS, Cell Counting, Immunohistochemistry, Marker, Staining

    Exogenous H 2 S alleviated inflammation response in ischemic myocardium from SOD1 KO mice. (a–b) Quantitative analysis of neutrophils (a) and macrophages (b) from SOD1 KO heart samples. (c) Immunohistochemistry of CD11b from SOD1 KO mice. Positive staining is shown in brown. Scale bar = 50 μ m. (d–e) Quantitative analysis of inflammatory cytokines IL-6 (d) and TNF- α (e) in heart tissues. (f–g) Quantitative analysis of neutrophils (f) and monocytes (g) in blood samples from SOD1 KO mice. Values are mean ± SE, n = 6 in each group. ∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hydrogen Sulfide Alleviates Acute Myocardial Ischemia Injury by Modulating Autophagy and Inflammation Response under Oxidative Stress

    doi: 10.1155/2018/3402809

    Figure Lengend Snippet: Exogenous H 2 S alleviated inflammation response in ischemic myocardium from SOD1 KO mice. (a–b) Quantitative analysis of neutrophils (a) and macrophages (b) from SOD1 KO heart samples. (c) Immunohistochemistry of CD11b from SOD1 KO mice. Positive staining is shown in brown. Scale bar = 50 μ m. (d–e) Quantitative analysis of inflammatory cytokines IL-6 (d) and TNF- α (e) in heart tissues. (f–g) Quantitative analysis of neutrophils (f) and monocytes (g) in blood samples from SOD1 KO mice. Values are mean ± SE, n = 6 in each group. ∗ p

    Article Snippet: IL-6 was detected using a rat anti-mouse IL-6 monoclonal antibody (504502, BioLegend, San Diego, CA, USA) for capture and biotinylated rat anti-mouse IL-6 monoclonal antibody (504602, BioLegend, San Diego, CA, USA) for detection.

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    SOD1 KO mice subjected to AMI showed severe inflammation response in the ischemic myocardium. (a) Representative FACS results of inflammatory cell counting from heart samples. (b–c) Quantitative analysis of neutrophils (b) and macrophages (c) in heart tissues. (d) Immunohistochemistry of CD11b (marker of both neutrophils and macrophages). Positive staining is shown in brown. Scale bar = 50 μ m. (e–f) Quantitative analysis of inflammatory cytokines IL-6 (e) and TNF- α (f) in heart tissues. (g–h) Quantitative analysis of neutrophils (g) and monocytes (h) in blood samples. Values are mean ± SE, n = 6 in each group. ∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hydrogen Sulfide Alleviates Acute Myocardial Ischemia Injury by Modulating Autophagy and Inflammation Response under Oxidative Stress

    doi: 10.1155/2018/3402809

    Figure Lengend Snippet: SOD1 KO mice subjected to AMI showed severe inflammation response in the ischemic myocardium. (a) Representative FACS results of inflammatory cell counting from heart samples. (b–c) Quantitative analysis of neutrophils (b) and macrophages (c) in heart tissues. (d) Immunohistochemistry of CD11b (marker of both neutrophils and macrophages). Positive staining is shown in brown. Scale bar = 50 μ m. (e–f) Quantitative analysis of inflammatory cytokines IL-6 (e) and TNF- α (f) in heart tissues. (g–h) Quantitative analysis of neutrophils (g) and monocytes (h) in blood samples. Values are mean ± SE, n = 6 in each group. ∗ p

    Article Snippet: IL-6 was detected using a rat anti-mouse IL-6 monoclonal antibody (504502, BioLegend, San Diego, CA, USA) for capture and biotinylated rat anti-mouse IL-6 monoclonal antibody (504602, BioLegend, San Diego, CA, USA) for detection.

    Techniques: Mouse Assay, FACS, Cell Counting, Immunohistochemistry, Marker, Staining

    The absence of secretory leukoprotease inhibitor (SLPI) in BM-derived eosinophils (BMEos) increases interleukin (IL)-6 production and invasive activity. (A) I The production of IL-6 by B6 and Slpi −/− BMEos after lipopolysaccharide (LPS) stimulation for 12 h. (B) B6 and Slpi −/− BMEos were incubated with LPS (1 µg/ml) or IL-33 (0.1 µg/ml) for 12 h. BMEos were also incubated with TNP-OVA (1 ng/ml) for 12 h after the administration of 5 µg/ml anti-TNP-IgE. (A,B) IL-4, 6, and 13 levels in the supernatants of cells were measured by an ELISA. (C) The activities of tryptase and chymase prepared according to the methods described in Figure 2 B. (D) The amounts of eosinophil peroxidase (EPO) in B6 and Slpi −/− BMEos after the administration of the indicated stimulators. (E) Chemotactic assays of B6 and Slpi −/− eosinophils by LTB 4 (50 nM), CCL2 (50 nM), and CCL11 (10 nM). (F) Invasion assays using Matrigel in B6 and Slpi −/− eosinophils upon LPS (1 µg/ml) and CCL11 (10 nM) stimulation. All of the data are shown as the mean ± SEM of three different eosinophil cultures. * P

    Journal: Frontiers in Immunology

    Article Title: Identification of Secretory Leukoprotease Inhibitor As an Endogenous Negative Regulator in Allergic Effector Cells

    doi: 10.3389/fimmu.2017.01538

    Figure Lengend Snippet: The absence of secretory leukoprotease inhibitor (SLPI) in BM-derived eosinophils (BMEos) increases interleukin (IL)-6 production and invasive activity. (A) I The production of IL-6 by B6 and Slpi −/− BMEos after lipopolysaccharide (LPS) stimulation for 12 h. (B) B6 and Slpi −/− BMEos were incubated with LPS (1 µg/ml) or IL-33 (0.1 µg/ml) for 12 h. BMEos were also incubated with TNP-OVA (1 ng/ml) for 12 h after the administration of 5 µg/ml anti-TNP-IgE. (A,B) IL-4, 6, and 13 levels in the supernatants of cells were measured by an ELISA. (C) The activities of tryptase and chymase prepared according to the methods described in Figure 2 B. (D) The amounts of eosinophil peroxidase (EPO) in B6 and Slpi −/− BMEos after the administration of the indicated stimulators. (E) Chemotactic assays of B6 and Slpi −/− eosinophils by LTB 4 (50 nM), CCL2 (50 nM), and CCL11 (10 nM). (F) Invasion assays using Matrigel in B6 and Slpi −/− eosinophils upon LPS (1 µg/ml) and CCL11 (10 nM) stimulation. All of the data are shown as the mean ± SEM of three different eosinophil cultures. * P

    Article Snippet: The mouse IL-4, IL-6 ELISA MAX Standard kit (Biolegend), the IL-13 Ready-Set-Go ELISA set (Thermo Fisher Scientific), a Histamine ELISA test kit (Neogen), and a Cysteinyl leukotrine ELISA kit (Cayman Chemical) were used.

    Techniques: Derivative Assay, Activity Assay, Incubation, Enzyme-linked Immunosorbent Assay

    C5a and C3a suppress IFN-β production in BMDCs WT BMDCs were pre-treated with either media, 50 nM C5a, or 500 nM C3a for 2 h, and then the cells were infected with Lm overnight. Cell-free supernatants were used to quantitate (A) IFN-β production or (B) IL-6, TNF-α, and MCP-1 production. (C) WT BMDCs were pre-treated with either media, 50 nM C5a, 100 nM C5aA, 500 nM C3a, or 100 nM C3aA for 2 h and then were incubated with 25 μg/ml c-di-AMP for 20 h. IFN-β was quantitated from cell-free supernatants. (D) C5aR−/− BMDCs were pre-treated with either media, 50 nM C5a, or 100 nM C5aA for 2 h and (E) C3aR−/− BMDCs were pre-treated with either media, 500 nM C3a, or 100 nM C3aA for 2 h, and then the cells were incubated with 25 μg/ml c-di-AMP for 20 h. IFN-β was quantitated from cell-free supernatants. All data are presented as mean pg/ml ± SEM. These data are pooled from three independent experiments. * P ≤ 0.017; ** P ≤ 0.009; *** P = 0.0006 by t test.

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

    Article Title: The Complement Anaphylatoxins, C5a and C3a, Suppress Interferon-Beta Production in Response to Listeria monocytogenes by Inhibition of the Cyclic Dinucleotide-Activated Cytosolic Surveillance Pathway

    doi: 10.4049/jimmunol.1601420

    Figure Lengend Snippet: C5a and C3a suppress IFN-β production in BMDCs WT BMDCs were pre-treated with either media, 50 nM C5a, or 500 nM C3a for 2 h, and then the cells were infected with Lm overnight. Cell-free supernatants were used to quantitate (A) IFN-β production or (B) IL-6, TNF-α, and MCP-1 production. (C) WT BMDCs were pre-treated with either media, 50 nM C5a, 100 nM C5aA, 500 nM C3a, or 100 nM C3aA for 2 h and then were incubated with 25 μg/ml c-di-AMP for 20 h. IFN-β was quantitated from cell-free supernatants. (D) C5aR−/− BMDCs were pre-treated with either media, 50 nM C5a, or 100 nM C5aA for 2 h and (E) C3aR−/− BMDCs were pre-treated with either media, 500 nM C3a, or 100 nM C3aA for 2 h, and then the cells were incubated with 25 μg/ml c-di-AMP for 20 h. IFN-β was quantitated from cell-free supernatants. All data are presented as mean pg/ml ± SEM. These data are pooled from three independent experiments. * P ≤ 0.017; ** P ≤ 0.009; *** P = 0.0006 by t test.

    Article Snippet: Mouse IL-6, TNF, and MCP-1 were quantitated in cell-free culture supernatants using ELISA MAX kits from BioLegend.

    Techniques: Infection, Incubation

    C5a and C3a suppress IFN-β production in J774A.1 cells J774A.1 cells were pre-treated with either media, 50 nM C5a, or 500 nM C3a for 2 h and then the cells were infected with Lm for 20 h. Cell-free supernatants were used to quantitate (A) IFN-β production or (B) IL-6, TNF-α, and MCP-1 production. J774A.1 cells were pre-treated with either media, 50 nM C5a, or 500 nM C3a for 2 h and then were incubated with (C) 25 μg/ml c-di-AMP or (D) 100 ng/ml LPS for 20 h. IFN-β was quantitated from cell-free supernatants. (E) J774A.1 cells were pre-treated with either media or 50 nM C5a for 30 min, 1 h, 2 h, 4 h, or 8 h and then were incubated with 100 ng/ml LPS for 20 h. IFN-β was quantitated from cell-free supernatants. All data are presented as mean pg/ml ± SEM. These data are pooled from three independent experiments. * P = 0.027; ** P ≤ 0.006; *** P ≤ 0.0001 by t test.

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

    Article Title: The Complement Anaphylatoxins, C5a and C3a, Suppress Interferon-Beta Production in Response to Listeria monocytogenes by Inhibition of the Cyclic Dinucleotide-Activated Cytosolic Surveillance Pathway

    doi: 10.4049/jimmunol.1601420

    Figure Lengend Snippet: C5a and C3a suppress IFN-β production in J774A.1 cells J774A.1 cells were pre-treated with either media, 50 nM C5a, or 500 nM C3a for 2 h and then the cells were infected with Lm for 20 h. Cell-free supernatants were used to quantitate (A) IFN-β production or (B) IL-6, TNF-α, and MCP-1 production. J774A.1 cells were pre-treated with either media, 50 nM C5a, or 500 nM C3a for 2 h and then were incubated with (C) 25 μg/ml c-di-AMP or (D) 100 ng/ml LPS for 20 h. IFN-β was quantitated from cell-free supernatants. (E) J774A.1 cells were pre-treated with either media or 50 nM C5a for 30 min, 1 h, 2 h, 4 h, or 8 h and then were incubated with 100 ng/ml LPS for 20 h. IFN-β was quantitated from cell-free supernatants. All data are presented as mean pg/ml ± SEM. These data are pooled from three independent experiments. * P = 0.027; ** P ≤ 0.006; *** P ≤ 0.0001 by t test.

    Article Snippet: Mouse IL-6, TNF, and MCP-1 were quantitated in cell-free culture supernatants using ELISA MAX kits from BioLegend.

    Techniques: Infection, Incubation

    The expression of IL-6 in cardiac mesenchymal fibroblasts is affected by aging. A ) Expression of IL-6 in quiescent fibroblasts derived from young and old MSCs were analyzed by quantitative PCR and by ELISA, 24 h after the cell cycle was synchronized. B ) Double immunofluorescence labeling of IL-6 (red) and DDR2 (green) in 3- and 30-mo-old mouse hearts. Double positive cells are visualized in yellow/orange color. Middle panel shows magnified image of the section marked by white squares. Bottom panel shows morphometric analysis of IL-6 + DDR2 + cells in heart sections. Nuclei were stained with DAPI. Scale bar, 20 μm. P

    Journal: The FASEB Journal

    Article Title: Mesenchymal stem cell-derived inflammatory fibroblasts promote monocyte transition into myeloid fibroblasts via an IL-6-dependent mechanism in the aging mouse heart

    doi: 10.1096/fj.14-268136

    Figure Lengend Snippet: The expression of IL-6 in cardiac mesenchymal fibroblasts is affected by aging. A ) Expression of IL-6 in quiescent fibroblasts derived from young and old MSCs were analyzed by quantitative PCR and by ELISA, 24 h after the cell cycle was synchronized. B ) Double immunofluorescence labeling of IL-6 (red) and DDR2 (green) in 3- and 30-mo-old mouse hearts. Double positive cells are visualized in yellow/orange color. Middle panel shows magnified image of the section marked by white squares. Bottom panel shows morphometric analysis of IL-6 + DDR2 + cells in heart sections. Nuclei were stained with DAPI. Scale bar, 20 μm. P

    Article Snippet: PD 0325901 was purchased from Cayman Chemical (Ann Arbor, MI, USA), Jak inhibitor was purchased from EMD Millipore (Billerica, MA, USA), FTI-277 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), brefeldin A was obtained from Sigma-Aldrich (St. Louis, MO, USA), mouse recombinant IL-6 was acquired from Biolegend (San Diego, CA, USA), and human recombinant MCP-1, human IL-6, soluble human IL-6R, soluble human gp130, and sc-144 hydrochloride were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Expressing, Derivative Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Labeling, Staining

    The FTase-ERK pathway contributes to IL-6 expression in cardiac fibroblasts. Quiescent fibroblasts derived from the old MSCs were treated with 100 nM FTI-277 (FTI) or 2 μ M PD 0325901 (PD) for 24 h. A–C ) IL-6 mRNA level ( A ), IL-6 secretion ( B ), and cytoplasmic IL-6 ( C ) are shown. To visualize cytoplasmic IL-6 cells were treated with 10 ng/ml brefeldin A for 6 h. Scale bar, 50 μm. P

    Journal: The FASEB Journal

    Article Title: Mesenchymal stem cell-derived inflammatory fibroblasts promote monocyte transition into myeloid fibroblasts via an IL-6-dependent mechanism in the aging mouse heart

    doi: 10.1096/fj.14-268136

    Figure Lengend Snippet: The FTase-ERK pathway contributes to IL-6 expression in cardiac fibroblasts. Quiescent fibroblasts derived from the old MSCs were treated with 100 nM FTI-277 (FTI) or 2 μ M PD 0325901 (PD) for 24 h. A–C ) IL-6 mRNA level ( A ), IL-6 secretion ( B ), and cytoplasmic IL-6 ( C ) are shown. To visualize cytoplasmic IL-6 cells were treated with 10 ng/ml brefeldin A for 6 h. Scale bar, 50 μm. P

    Article Snippet: PD 0325901 was purchased from Cayman Chemical (Ann Arbor, MI, USA), Jak inhibitor was purchased from EMD Millipore (Billerica, MA, USA), FTI-277 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), brefeldin A was obtained from Sigma-Aldrich (St. Louis, MO, USA), mouse recombinant IL-6 was acquired from Biolegend (San Diego, CA, USA), and human recombinant MCP-1, human IL-6, soluble human IL-6R, soluble human gp130, and sc-144 hydrochloride were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Expressing, Derivative Assay

    Differential response to recombinant IL-6 in fibroblasts derived from young and old MSCs. Quiescent fibroblasts were treated with one dose of recombinant IL-6 (50 ng/ml) and then cells were harvested 24 and 48 h later. A ) Response of fibroblast derived from the young MSCs. B ) Responses from fibroblasts derived from the old MSCs. P

    Journal: The FASEB Journal

    Article Title: Mesenchymal stem cell-derived inflammatory fibroblasts promote monocyte transition into myeloid fibroblasts via an IL-6-dependent mechanism in the aging mouse heart

    doi: 10.1096/fj.14-268136

    Figure Lengend Snippet: Differential response to recombinant IL-6 in fibroblasts derived from young and old MSCs. Quiescent fibroblasts were treated with one dose of recombinant IL-6 (50 ng/ml) and then cells were harvested 24 and 48 h later. A ) Response of fibroblast derived from the young MSCs. B ) Responses from fibroblasts derived from the old MSCs. P

    Article Snippet: PD 0325901 was purchased from Cayman Chemical (Ann Arbor, MI, USA), Jak inhibitor was purchased from EMD Millipore (Billerica, MA, USA), FTI-277 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), brefeldin A was obtained from Sigma-Aldrich (St. Louis, MO, USA), mouse recombinant IL-6 was acquired from Biolegend (San Diego, CA, USA), and human recombinant MCP-1, human IL-6, soluble human IL-6R, soluble human gp130, and sc-144 hydrochloride were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Recombinant, Derivative Assay

    RasGrf1 expression controls IL-6 transcription in fibroblasts derived from the old MSCs. A ). Silencing RasGrf1 by siRNA in fibroblasts derived from old MSCs reduced RasGrf1 mRNA (left panel) and protein (right panel) levels. Fibroblasts were transfected with scrambled siRNA (labeled: control) or siRNA targeting RasGrf1 (labeled: siRNA). RNA and protein analysis was carried 48 h posttransfection. B and C ) RasGrf1 silencing caused a decrease of IL-6 transcription ( B ) and reduced levels of secreted IL-6 ( C ). D) Cells transfected with scrambled siRNA or RasGrf1 siRNA were treated with 10 ng/ml brefeldin A for 6 h (starting 42 h posttransfection). Cytoplasmic IL-6 was visualized by immunofluorescence labeling. Scale bar, 50 μm. P

    Journal: The FASEB Journal

    Article Title: Mesenchymal stem cell-derived inflammatory fibroblasts promote monocyte transition into myeloid fibroblasts via an IL-6-dependent mechanism in the aging mouse heart

    doi: 10.1096/fj.14-268136

    Figure Lengend Snippet: RasGrf1 expression controls IL-6 transcription in fibroblasts derived from the old MSCs. A ). Silencing RasGrf1 by siRNA in fibroblasts derived from old MSCs reduced RasGrf1 mRNA (left panel) and protein (right panel) levels. Fibroblasts were transfected with scrambled siRNA (labeled: control) or siRNA targeting RasGrf1 (labeled: siRNA). RNA and protein analysis was carried 48 h posttransfection. B and C ) RasGrf1 silencing caused a decrease of IL-6 transcription ( B ) and reduced levels of secreted IL-6 ( C ). D) Cells transfected with scrambled siRNA or RasGrf1 siRNA were treated with 10 ng/ml brefeldin A for 6 h (starting 42 h posttransfection). Cytoplasmic IL-6 was visualized by immunofluorescence labeling. Scale bar, 50 μm. P

    Article Snippet: PD 0325901 was purchased from Cayman Chemical (Ann Arbor, MI, USA), Jak inhibitor was purchased from EMD Millipore (Billerica, MA, USA), FTI-277 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), brefeldin A was obtained from Sigma-Aldrich (St. Louis, MO, USA), mouse recombinant IL-6 was acquired from Biolegend (San Diego, CA, USA), and human recombinant MCP-1, human IL-6, soluble human IL-6R, soluble human gp130, and sc-144 hydrochloride were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Expressing, Derivative Assay, Transfection, Labeling, Immunofluorescence

    Role of elevated RasGrf1 expression in amplified IL-6 levels in fibroblasts derived from the old MSCs. Increased expression of RasGrf1 promotes exchange of GDP into GTP on Ras and renders its activation. Ras activation is also controlled by FTase activity since use of the FTase inhibitor (FTI-277) reduces IL-6 expression. Downstream of Ras and FTase, ERK activation controls IL-6 expression; use of PD 0325901 (ERK inhibitor) down-regulates IL-6 levels. Finally, IL-6 that is secreted activates its own transcription in a feed-forward loop.

    Journal: The FASEB Journal

    Article Title: Mesenchymal stem cell-derived inflammatory fibroblasts promote monocyte transition into myeloid fibroblasts via an IL-6-dependent mechanism in the aging mouse heart

    doi: 10.1096/fj.14-268136

    Figure Lengend Snippet: Role of elevated RasGrf1 expression in amplified IL-6 levels in fibroblasts derived from the old MSCs. Increased expression of RasGrf1 promotes exchange of GDP into GTP on Ras and renders its activation. Ras activation is also controlled by FTase activity since use of the FTase inhibitor (FTI-277) reduces IL-6 expression. Downstream of Ras and FTase, ERK activation controls IL-6 expression; use of PD 0325901 (ERK inhibitor) down-regulates IL-6 levels. Finally, IL-6 that is secreted activates its own transcription in a feed-forward loop.

    Article Snippet: PD 0325901 was purchased from Cayman Chemical (Ann Arbor, MI, USA), Jak inhibitor was purchased from EMD Millipore (Billerica, MA, USA), FTI-277 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), brefeldin A was obtained from Sigma-Aldrich (St. Louis, MO, USA), mouse recombinant IL-6 was acquired from Biolegend (San Diego, CA, USA), and human recombinant MCP-1, human IL-6, soluble human IL-6R, soluble human gp130, and sc-144 hydrochloride were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Expressing, Amplification, Derivative Assay, Activation Assay, Activity Assay

    IL-6 facilitates MCP-1-driven monocyte to myeloid fibroblast maturation. For experiments ( A–C ) PBMCs were added on top of a confluent endothelial cell monolayer seeded on an insert. Medium below the insert was supplemented as indicated. The following reagents were used: IL-6 (10 ng/ml), MCP-1 (650 ng/ml), IL-6R (200 ng/ml), soluble gp130 (300 ng/ml), sc-144 hydrochloride (5 μM), PD 0325901 (1 μM), Jak inhibitor (2 μM). noRX, no supplementation; sc-144, sc-144 hydrochloride; PD, PD 0325901; Jaki, Jak inhibitor. D ) Fibroblasts derived from young and old MSCs secrete the same level of soluble IL-6R. P

    Journal: The FASEB Journal

    Article Title: Mesenchymal stem cell-derived inflammatory fibroblasts promote monocyte transition into myeloid fibroblasts via an IL-6-dependent mechanism in the aging mouse heart

    doi: 10.1096/fj.14-268136

    Figure Lengend Snippet: IL-6 facilitates MCP-1-driven monocyte to myeloid fibroblast maturation. For experiments ( A–C ) PBMCs were added on top of a confluent endothelial cell monolayer seeded on an insert. Medium below the insert was supplemented as indicated. The following reagents were used: IL-6 (10 ng/ml), MCP-1 (650 ng/ml), IL-6R (200 ng/ml), soluble gp130 (300 ng/ml), sc-144 hydrochloride (5 μM), PD 0325901 (1 μM), Jak inhibitor (2 μM). noRX, no supplementation; sc-144, sc-144 hydrochloride; PD, PD 0325901; Jaki, Jak inhibitor. D ) Fibroblasts derived from young and old MSCs secrete the same level of soluble IL-6R. P

    Article Snippet: PD 0325901 was purchased from Cayman Chemical (Ann Arbor, MI, USA), Jak inhibitor was purchased from EMD Millipore (Billerica, MA, USA), FTI-277 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), brefeldin A was obtained from Sigma-Aldrich (St. Louis, MO, USA), mouse recombinant IL-6 was acquired from Biolegend (San Diego, CA, USA), and human recombinant MCP-1, human IL-6, soluble human IL-6R, soluble human gp130, and sc-144 hydrochloride were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Derivative Assay

    JAK1 interacts Sirt1. A , JAK1 expression plasmid was co-transfected with or without FLAG-Sirt1 into HEK293 cells. The interaction of Sirt1 with JAK1 in the transfected cells was analyzed by co-immunoprecipitation with anti-FLAG antibody and by Western blotting with JAK1 antibody ( top panel ). The same membrane was reprobed with anti-Sirt1 ( middle panel ). JAK1 expression in the whole cell lysate was determined by Western blotting ( bottom panel ). B–D , MCF-10 ( B ), HCT116 ( C ), and MM.1s ( D ) cells were stimulated with or without 10 ng/ml IL-6 and lysed with RIPA buffer. The interaction of Sirt1 and JAK1 was determined by immunoprecipitation of JAK1 using normal rabbit IgG ( rIgG ) as a control and by Western blotting with anti-Sirt1 antibody ( top panels ). The same membrane was stripped and reblotted with anti-JAK1 ( middle panels ). Sirt1 expression in the whole cell lysate was determined by Western blotting ( bottom panels ).

    Journal: The Journal of Biological Chemistry

    Article Title: JAK1-mediated Sirt1 phosphorylation functions as a negative feedback of the JAK1-STAT3 pathway

    doi: 10.1074/jbc.RA117.001387

    Figure Lengend Snippet: JAK1 interacts Sirt1. A , JAK1 expression plasmid was co-transfected with or without FLAG-Sirt1 into HEK293 cells. The interaction of Sirt1 with JAK1 in the transfected cells was analyzed by co-immunoprecipitation with anti-FLAG antibody and by Western blotting with JAK1 antibody ( top panel ). The same membrane was reprobed with anti-Sirt1 ( middle panel ). JAK1 expression in the whole cell lysate was determined by Western blotting ( bottom panel ). B–D , MCF-10 ( B ), HCT116 ( C ), and MM.1s ( D ) cells were stimulated with or without 10 ng/ml IL-6 and lysed with RIPA buffer. The interaction of Sirt1 and JAK1 was determined by immunoprecipitation of JAK1 using normal rabbit IgG ( rIgG ) as a control and by Western blotting with anti-Sirt1 antibody ( top panels ). The same membrane was stripped and reblotted with anti-JAK1 ( middle panels ). Sirt1 expression in the whole cell lysate was determined by Western blotting ( bottom panels ).

    Article Snippet: Human recombinant IL-6 was purchased from BioLegend (San Diego, CA) and the JAK1 inhibitor was from Sigma-Aldrich.

    Techniques: Expressing, Plasmid Preparation, Transfection, Immunoprecipitation, Western Blot

    JAK1-mediated Sirt1 phosphorylation inhibits STAT3 transcription activity. A , MCF-10 cells were transfected with STAT3 and FLAG-Sirt1 or its YF mutant. STAT3 acetylation was determined by immunoprecipitation with anti-STAT3 and by Western blotting with anti–acetyl-lysine antibody ( top panel ). The same membrane was reprobed with anti-STAT3 ( middle panel ), and the expression levels of Sirt1 in the whole cell lysate were analyzed by Western blotting ( bottom panel ). B and C , MCF-10 cells were transfected with Sirt1 or its YF mutant. The transfected cells were stimulated with IL-6 for 6 h and the binding of STAT3 ( B ) and Sirt1 ( C ) onto Bcl2 or Mcl1 promoter was determined by ChIP. D , MCF-10 cells were treated with IL-6 or together with JAK1 inhibitor PF-04965842; the binding of Sirt1 onto Bcl2 or Mcl1 promoter was determined by ChIP. E and F , MCF-10 cells were transfected with Sirt1 or its YF mutant. The transfected cells were stimulated with IL-6 for 6 h and the levels of H3K8Ac ( E ) and H4K16Ac ( F ) at the Bcl2 or Mcl1 promoter was determined by ChIP. The Student's t test was used for the statistical analysis. NS , not significantly different; *, p

    Journal: The Journal of Biological Chemistry

    Article Title: JAK1-mediated Sirt1 phosphorylation functions as a negative feedback of the JAK1-STAT3 pathway

    doi: 10.1074/jbc.RA117.001387

    Figure Lengend Snippet: JAK1-mediated Sirt1 phosphorylation inhibits STAT3 transcription activity. A , MCF-10 cells were transfected with STAT3 and FLAG-Sirt1 or its YF mutant. STAT3 acetylation was determined by immunoprecipitation with anti-STAT3 and by Western blotting with anti–acetyl-lysine antibody ( top panel ). The same membrane was reprobed with anti-STAT3 ( middle panel ), and the expression levels of Sirt1 in the whole cell lysate were analyzed by Western blotting ( bottom panel ). B and C , MCF-10 cells were transfected with Sirt1 or its YF mutant. The transfected cells were stimulated with IL-6 for 6 h and the binding of STAT3 ( B ) and Sirt1 ( C ) onto Bcl2 or Mcl1 promoter was determined by ChIP. D , MCF-10 cells were treated with IL-6 or together with JAK1 inhibitor PF-04965842; the binding of Sirt1 onto Bcl2 or Mcl1 promoter was determined by ChIP. E and F , MCF-10 cells were transfected with Sirt1 or its YF mutant. The transfected cells were stimulated with IL-6 for 6 h and the levels of H3K8Ac ( E ) and H4K16Ac ( F ) at the Bcl2 or Mcl1 promoter was determined by ChIP. The Student's t test was used for the statistical analysis. NS , not significantly different; *, p

    Article Snippet: Human recombinant IL-6 was purchased from BioLegend (San Diego, CA) and the JAK1 inhibitor was from Sigma-Aldrich.

    Techniques: Activity Assay, Transfection, Mutagenesis, Immunoprecipitation, Western Blot, Expressing, Binding Assay, Chromatin Immunoprecipitation

    B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and IL-6 ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P

    Journal: Scientific Reports

    Article Title: Thioacetamide-induced liver damage and thrombocytopenia is associated with induction of antiplatelet autoantibody in mice

    doi: 10.1038/s41598-019-53977-7

    Figure Lengend Snippet: B cell deficient (BCD) mice displayed markedly less liver damage, anti-PLT Ig, thrombocytopenia and TNF expression versus wild type mice. TAA-mediated induction of circulating AST ( A ), ALT ( B ), anti-PLT Ig ( C ; WT Day 0 groups were normalized to 1 fold), PLT counts ( D ), TNF-α ( E ), HMGB1 ( F ), and IL-6 ( G ) levels in B cell deficient (BCD) vs. wild type (WT) mice were shown. n = 6, # P

    Article Snippet: Rat anti mouse IL-6 antibody was purchased from BioLegend. (San Diego, CA).

    Techniques: Mouse Assay, Expressing, AST Assay

    Pml regulates pro-inflammatory cytokines in MSCs. a ELISA array for the detection of soluble factors and cytokines in the supernatant of MSCs Pml +/+ or Pml −/− . The quantification of significantly different factors is shown in the lower chart. b Relative expression of cxcl1 (charts on the left) and Il6 (charts on the right) in MSCs derived from Pml +/+ or Pml −/− mice (upper charts) or from Prrx1-Cre-Pml +/+ and Prrx1-Cre-Pml F /F mice (charts at the bottom). Cells of n = 3 mice each group have been pooled before RNA extraction. Data are shown as average ± SEM. c Western blot showing the reduction of Pml expression in HS5 cells treated with two independent shRNAs against Pml is shown on the left; the expression levels of cxcl1 and Il6 in cells silenced for the expression of Pml is shown in the charts on the right. Data are shown as average ± SEM

    Journal: Nature Communications

    Article Title: A non-cell-autonomous role for Pml in the maintenance of leukemia from the niche

    doi: 10.1038/s41467-017-02427-x

    Figure Lengend Snippet: Pml regulates pro-inflammatory cytokines in MSCs. a ELISA array for the detection of soluble factors and cytokines in the supernatant of MSCs Pml +/+ or Pml −/− . The quantification of significantly different factors is shown in the lower chart. b Relative expression of cxcl1 (charts on the left) and Il6 (charts on the right) in MSCs derived from Pml +/+ or Pml −/− mice (upper charts) or from Prrx1-Cre-Pml +/+ and Prrx1-Cre-Pml F /F mice (charts at the bottom). Cells of n = 3 mice each group have been pooled before RNA extraction. Data are shown as average ± SEM. c Western blot showing the reduction of Pml expression in HS5 cells treated with two independent shRNAs against Pml is shown on the left; the expression levels of cxcl1 and Il6 in cells silenced for the expression of Pml is shown in the charts on the right. Data are shown as average ± SEM

    Article Snippet: In selected experiments neutralizing antibodies anti-CXCR2 (R & D) (250 ng/ml) and anti-IL6R (Biolegend) (100 ng/ml), or the recombinant proteins CXCL1 and IL6 (both from Biolegend and added at the concentration of 100 ng/ml) were added to the co-cultures.

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Derivative Assay, Mouse Assay, RNA Extraction, Western Blot

    Pml regulates LICs non-cell autonomously through Il6 and Cxcl1 pathways. a The schematic overview of the experimental design is depicted in the upper panel. The numbers of leukemic cells with different genotype, derived from co-cultures treated with the combination of anti-IL6R and anti-CXCR2, are shown in the lower panel. Data are shown as average ± SEM. ( n ≥ 2 independent experiments). b Co-cultures of MSCs Pml +/+ and HoxA9–Meis1 + GFP + leukemic cells were untreated (NT) as control, or treated with anti-CXCR2 in combination of anti-IL6R. Co-cultures with MSCs Pml −/− and HoxA9–Meis1 + GFP + leukemic cells were untreated as control, or treated with recombinant Il6 and Cxcl1 proteins. Leukemic cells were then re-plated onto new MSCs ( Pml +/+ or Pml −/− ); secondary co-cultures were analyzed for the presence of GFP + ckit + cells. The relative numbers of GFP + ckit + cells in the different conditions are shown. Data are shown as average ± SEM. c Schematic representation of the proposed model. In MSCs, Pml regulates the secretion of pro-inflammatory soluble factors, which sustain the maintenance of leukemic cells and those enriched for LIC capacity. Pml sustains leukemic cells in a non-cell-autonomous manner, in cooperation with the cell-autonomous mechanisms within LICs, as previously described. d Pml degradation, (achieved for example through treatment with arsenic trioxide (AS 2 O 3 )), contributes to eliminate leukemic cells

    Journal: Nature Communications

    Article Title: A non-cell-autonomous role for Pml in the maintenance of leukemia from the niche

    doi: 10.1038/s41467-017-02427-x

    Figure Lengend Snippet: Pml regulates LICs non-cell autonomously through Il6 and Cxcl1 pathways. a The schematic overview of the experimental design is depicted in the upper panel. The numbers of leukemic cells with different genotype, derived from co-cultures treated with the combination of anti-IL6R and anti-CXCR2, are shown in the lower panel. Data are shown as average ± SEM. ( n ≥ 2 independent experiments). b Co-cultures of MSCs Pml +/+ and HoxA9–Meis1 + GFP + leukemic cells were untreated (NT) as control, or treated with anti-CXCR2 in combination of anti-IL6R. Co-cultures with MSCs Pml −/− and HoxA9–Meis1 + GFP + leukemic cells were untreated as control, or treated with recombinant Il6 and Cxcl1 proteins. Leukemic cells were then re-plated onto new MSCs ( Pml +/+ or Pml −/− ); secondary co-cultures were analyzed for the presence of GFP + ckit + cells. The relative numbers of GFP + ckit + cells in the different conditions are shown. Data are shown as average ± SEM. c Schematic representation of the proposed model. In MSCs, Pml regulates the secretion of pro-inflammatory soluble factors, which sustain the maintenance of leukemic cells and those enriched for LIC capacity. Pml sustains leukemic cells in a non-cell-autonomous manner, in cooperation with the cell-autonomous mechanisms within LICs, as previously described. d Pml degradation, (achieved for example through treatment with arsenic trioxide (AS 2 O 3 )), contributes to eliminate leukemic cells

    Article Snippet: In selected experiments neutralizing antibodies anti-CXCR2 (R & D) (250 ng/ml) and anti-IL6R (Biolegend) (100 ng/ml), or the recombinant proteins CXCL1 and IL6 (both from Biolegend and added at the concentration of 100 ng/ml) were added to the co-cultures.

    Techniques: Derivative Assay, Recombinant

    iSN34 with CpG-B enhances IL-6 production. Supernatants from stimulated cells were collected and IL-6 protein levels were measured by ELISA. Mouse splenocytes were harvested 48 h later and intracellular IL-6 protein levels were determined by ELISA. All assays were carried out at least three independent times in triplicate. Similar results were obtained from at least three different mice. Values are presented as mean + SD of three independent experiments, each performed in triplicate ( n = 9). Values with different letters (i.e., a, b, c, d, e, and f) were significantly different. **** p

    Journal: BMC Immunology

    Article Title: Synergistic oligodeoxynucleotide strongly promotes CpG-induced interleukin-6 production

    doi: 10.1186/s12865-017-0227-7

    Figure Lengend Snippet: iSN34 with CpG-B enhances IL-6 production. Supernatants from stimulated cells were collected and IL-6 protein levels were measured by ELISA. Mouse splenocytes were harvested 48 h later and intracellular IL-6 protein levels were determined by ELISA. All assays were carried out at least three independent times in triplicate. Similar results were obtained from at least three different mice. Values are presented as mean + SD of three independent experiments, each performed in triplicate ( n = 9). Values with different letters (i.e., a, b, c, d, e, and f) were significantly different. **** p

    Article Snippet: For intracellular staining, cells were fixed in 4% PFA for 15 min at room temperature, washed, and permeabilized by incubation for 15 min on ice; cells were then further incubated with phycoerythrin-labeled anti-mouse IL-6 antibody (Biolegend).

    Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay

    Representative flow cytometry plots. Dot plot of forward-angle versus right-angle light-scattering properties; the oval marks indicate the electronic windows used for analysis of fluorescence data for lymphocyte precursors; the rectangular boxes indicate the electronic windows used for analysis of fluorescence data for monocyte precursors with percentages. a A quadrant has been set to delineate the CD19 and IL-6 cells. Murine splenocytes were stimulated with water + water, water + CpG-B, iSN34 + water, or iSN34 + CpG-B for 12 h, and then sorted into CD19 + IL-6 + cells. b Mean percentage of IL-6 + CD19 + cells in the total population was determined in each group. Similar results were obtained from at least three different mice. Values are presented as mean + SD of three independent experiments, each performed in triplicate ( n = 9). Values with different letters (i.e., a, b, c, and d) were significantly different ( p

    Journal: BMC Immunology

    Article Title: Synergistic oligodeoxynucleotide strongly promotes CpG-induced interleukin-6 production

    doi: 10.1186/s12865-017-0227-7

    Figure Lengend Snippet: Representative flow cytometry plots. Dot plot of forward-angle versus right-angle light-scattering properties; the oval marks indicate the electronic windows used for analysis of fluorescence data for lymphocyte precursors; the rectangular boxes indicate the electronic windows used for analysis of fluorescence data for monocyte precursors with percentages. a A quadrant has been set to delineate the CD19 and IL-6 cells. Murine splenocytes were stimulated with water + water, water + CpG-B, iSN34 + water, or iSN34 + CpG-B for 12 h, and then sorted into CD19 + IL-6 + cells. b Mean percentage of IL-6 + CD19 + cells in the total population was determined in each group. Similar results were obtained from at least three different mice. Values are presented as mean + SD of three independent experiments, each performed in triplicate ( n = 9). Values with different letters (i.e., a, b, c, and d) were significantly different ( p

    Article Snippet: For intracellular staining, cells were fixed in 4% PFA for 15 min at room temperature, washed, and permeabilized by incubation for 15 min on ice; cells were then further incubated with phycoerythrin-labeled anti-mouse IL-6 antibody (Biolegend).

    Techniques: Flow Cytometry, Cytometry, Fluorescence, Mouse Assay

    iSN34 was used to determine the optimal concentration of ODNs. a Mouse splenocytes were pre-incubated in medium for 3 h prior to exposure to iSN34 (at 0.01, 0.04, 0.16, 0.63, 2.5, or 10 μM) + CpG-B (ODN1555; at equimolar levels) or to ODN 1612 (control) for 6 h. Accumulation of IL-6 mRNA was determined by qPCR. Results are shown as the ratio of IL-6 mRNA levels for stimulated (iSN + CpG-B) versus ODN 1612-treated cells. b The synergistic effects of iSN34 (0.63 μM) were assessed in combination with CpG-A (ODN 1585), CpG-B (ODN 1555), CpG-C (ODN 2395), and Ctr 1612 (Control ODN). c Mouse splenocytes (1 × 10 7 cells/mL) were pre-incubated in medium for 3 h prior to exposure to 3 μM ODN 1612 or iSN34 for 24 h. Cells then were washed with medium (to remove the ODNs) and resuspended in medium with 3 μM CpG-B (ODN 1555) for 6 h. Results are shown as IL-6 mRNA expression (normalized to β-actin-encoding mRNA; see qPCR method) in stimulated cells in the wash-out assay. All assays were carried out at least three independent times in triplicate. Similar results were obtained from at least three different mice. Values are presented as mean + SD of three independent experiments, each performed in triplicate ( n = 9). Values with different letters (i.e. , a, b, c, d, and e) were significantly different. **** p

    Journal: BMC Immunology

    Article Title: Synergistic oligodeoxynucleotide strongly promotes CpG-induced interleukin-6 production

    doi: 10.1186/s12865-017-0227-7

    Figure Lengend Snippet: iSN34 was used to determine the optimal concentration of ODNs. a Mouse splenocytes were pre-incubated in medium for 3 h prior to exposure to iSN34 (at 0.01, 0.04, 0.16, 0.63, 2.5, or 10 μM) + CpG-B (ODN1555; at equimolar levels) or to ODN 1612 (control) for 6 h. Accumulation of IL-6 mRNA was determined by qPCR. Results are shown as the ratio of IL-6 mRNA levels for stimulated (iSN + CpG-B) versus ODN 1612-treated cells. b The synergistic effects of iSN34 (0.63 μM) were assessed in combination with CpG-A (ODN 1585), CpG-B (ODN 1555), CpG-C (ODN 2395), and Ctr 1612 (Control ODN). c Mouse splenocytes (1 × 10 7 cells/mL) were pre-incubated in medium for 3 h prior to exposure to 3 μM ODN 1612 or iSN34 for 24 h. Cells then were washed with medium (to remove the ODNs) and resuspended in medium with 3 μM CpG-B (ODN 1555) for 6 h. Results are shown as IL-6 mRNA expression (normalized to β-actin-encoding mRNA; see qPCR method) in stimulated cells in the wash-out assay. All assays were carried out at least three independent times in triplicate. Similar results were obtained from at least three different mice. Values are presented as mean + SD of three independent experiments, each performed in triplicate ( n = 9). Values with different letters (i.e. , a, b, c, d, and e) were significantly different. **** p

    Article Snippet: For intracellular staining, cells were fixed in 4% PFA for 15 min at room temperature, washed, and permeabilized by incubation for 15 min on ice; cells were then further incubated with phycoerythrin-labeled anti-mouse IL-6 antibody (Biolegend).

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