interleukin-6 Search Results


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  • 99
    Thermo Fisher interleukin 6
    Secretion of <t>interleukin-6</t> (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) from BM-MSCs after stimulation with sEV-OCS or sEV-OCS-Cis. ( A – C ) Secretion levels of ( A ) IL-6, ( B ) IL-8, and ( C ) VEGFA of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis measured by multiplex fluorescent bead-based immunoassay analysis. BM-MSCs cell-cultured media (CM) was harvested 24 hours post-stimulation with sEV-OCS (CM MCS -sEV-OCS) or sEV-OCS-Cis (CM MCS -sEV-OCS-Cis). CM of BM-MSCs without sEV stimulation (CM MCS -non-sEV) was used as control. Results are mean ± SEM of n = 3. Statistical analysis was performed using the Kruskal–Wallis test followed by the Mann–Whitney test. * p
    Interleukin 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1698 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore interleukin 6
    Concentrations of ( a ) IL-1β, ( b ) <t>IL-6,</t> ( c ) TNF-α ( d ) anti-OVA Ab in serum determined by ELISA. The results are representative of means ± SD ( n = 5; * p
    Interleukin 6, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 625 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human il 6
    JSI-124 inhibited bleomycin-induced bronchoalveolar inflammatory cell extravasation and lung fibrosis. Wistar rats were administered a single intratracheal dose of bleomycin (BLM; 3.75 U/kg) on day 1. JSI-124 (1 mg/kg/day) or vehicle was administered intraperitoneally from day 14 until the analysis at day 28 ( n = 10 per group). Total bronchoalveolar lavage ( a ) protein, ( b , c ) inflammatory cells, ( d ) lung weight, ( e ) protein, and ( f , g ) <t>IL-6</t> and IL-13 content were measured at day 28. h Masson’s trichrome (upper panels, scale bar: 100 μm) of control, BLM, and BLM + JSI-124 tissue. i Fibrosis Ashcroft scores were assessed as described in the Methods. j Micro-CT images were acquired on day 28 and ( k ) quantified as Hounsfield units. The results are expressed as the mean ± SEM, n = 10. Statistical significance was assessed using a t -test or one-way ANOVA followed by a Bonferroni post-hoc test. * P
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    Becton Dickinson interleukin 6
    LPS-Trap-Fc1 to -Fc4 bind LPS and block LPS-mediated <t>IL-6</t> production in Mono Mac 6 cells. (A) Supernatants of HEK293T cells transfected with LPS-Trap-Fc1 were incubated in the presence (lane 1) or absence (lane 3) of biotin-LPS. Additionally, supernatants were preincubated with an anti-TLR4/MD-2 MAb (lane 2) or an excess of LPS (lane 4). Complexes were immunoprecipitated with streptavidin-Sepharose, subjected to SDS-PAGE, and analyzed by Western blotting with an anti-FLAG MAb. (B) RAW 264.7 cells were incubated with LPS-Trap-Fc1 or a medium control. Cells were stimulated with 100 ng/ml LPS overnight, and IL-6 levels in supernatants were determined by ELISA. *, P
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    R&D Systems mouse il 6
    <t>IL-6</t> is up-regulated in PDGFRα + stromal cells after SO 2 injury. ( A ) RNAs were extracted from whole trachea at 0, 1, 2, and 14 d after injury and subjected to quantitative RT-PCR analysis. The mRNA expression level of cytokines was normalized to Gapdh . ( B ) In situ hybridization combined with immunohistochemistry shows that Il-6 mRNA (red) is expressed in cells in the stroma beneath basal cells (K5 + , green) after SO 2 injury. ( C ) Quantitative PCR analysis of Il-6 expression in sorted stromal cells [ Pdgfr α ( Pdgfra )-GFP + ] and immune cell subpopulations from the trachea at 24 hpi. ( D ) Immunohistochemistry of a trachea section at 24 hpi shows Pdgfra -GFP + cells (GFP + , green) in the stroma beneath the epithelium with basal cells (K5 + , red). ( E ) In situ hybridization and immunohistochemistry show that Pdgfra -GFP + cells (GFP + , green) express Il-6 mRNA (red) at 24 hpi. (Scale bars: B and E , 20 μm; D , 50 μm.) * P
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    R&D Systems interleukin 6
    Niphatenone B enantiomers block transactivation of AR NTD. (A) Chemical structures of sintokamide, EPI-001, and niphatenone B enantiomers. (B) Transactivation assay of the AR NTD performed in LNCaP cells that were co-transfected with Gal4UAS-TATA-luciferase and AR-(1-558)-Gal4 DBD prior pre-treatment for 1 hour with 25 µM EPI-002, 7.0 µM niphatenones or DMSO vehicle control. Transactivation of AR NTD was induced by incubation with <t>IL-6</t> (50 ng/ml) for 24 hours. Luciferase activity was normalized to protein concentration. (C) Niphatenones inhibit the constitutively active AR V567es splice variant. Cos-1 cells co-transfected with PB-luciferase and an expression vector for ARvar567 were treated with each niphatenone B (1 µM) or DMSO vehicle control. After 24 hours of exposure, cells were harvested and luciferase activities were normalized to protein concentrations of the samples. Data represent the mean ± SEM of n = 3 separate experiments with triplicate wells. Student's t test: ** P
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    R&D Systems recombinant human il 6
    MCR expressions during erythroblast differentiation. ( A ) Schematic diagram of an in vitro differentiation protocol for deriving erythroblasts from human HPCs. Human CD34 + HPCs were expanded for 7 days (E0–E7) and stocks were frozen in liquid nitrogen (LN 2 ). The stock cells were differentiated for 3 days (D0–D3) before undergoing maturation (M0–M7). The number of enucleated erythrocytes increased from M5 to M7. ACTH, adrenocorticotropic hormone; EPO, erythropoietin; FL, flt-3 ligand; IL-3, interleukin-3; <t>IL-6,</t> interleukin-6; α-MSH, α-melanocyte stimulating hormone; SCF, stem cell factor; TPO, thrombopoietin. ( B ) May-Grunwald-Giemsa staining of control erythroblasts at day M0–5 day. Cells differentiated to Pro-E stage at day M0 and Baso-E stage at day M3. Arrows, Poly-E; Arrow heads, Orho-E. *, Reticulocyte. Bar 10 μm. ( C ) Conventional RT-PCR for MCRs during erythroblasts differentiation between M0 and M6 day.
    Recombinant Human Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 996 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher mouse il 6
    Biochemical and hematological parameters of DSS-induced colitis: the colonic levels of a MPO activity, b tGSH, c <t>IL-6</t> and d TNF-α; the serum levels of e IL-6 and f TNF-α; the hematologic results of (g) white blood cells. Each point represents a single sample. Asterisks and plus signs represent significant differences (*** P
    Mouse Il 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 553 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech recombinant mouse il 6
    WT-CCN1, but not DM-CCN1, induces <t>IL-6</t> in macrophages and fibroblasts (A) IL-6 mRNA was measured by qRT-PCR and (B) IL-6 protein from conditioned media was measured by ELISA in serum-starved (overnight) I13.35 macrophages treated with 5 μg/ml of purified WT-CCN1 or DM-CCN1, or BSA for 24 hrs. (C) I13.35 macrophages were incubated with blocking mAbs against integrin α M (50 μg/ml) or β 2 (50 μg/ml) 1 hr prior to treatment with WT-CCN1 for 24 hrs. IL-6 mRNA was measured by qRT-PCR. (D) IL-6 mRNA was measured by qRT-PCR and (E) IL-6 protein from conditioned media was measured by ELISA in serum-starved 18Co fibroblasts treated with 5 μg/ml of purified WT-CCN1, DM-CCN1, BSA, or 25 ng/ml of TNFα for 24 hrs. (F) 18Co fibroblasts were incubated with blocking mAb against integrin α 6 for 1 hr, then treated with WT-CCN1, TNFα, or BSA for 24 hrs as above. IL-6 mRNA was measured by qRT-PCR. Data shown as mean ± SD of triplicate experiments.
    Recombinant Mouse Il 6, supplied by PeproTech, used in various techniques. Bioz Stars score: 97/100, based on 435 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam anti il 6
    Comparison of TNF- α and <t>IL-6</t> production in the sciatic nerve after CCI and BGJTD/BeD treatments. Western blot analysis of TNF- α (a) and IL-6 (b) in the proximal (P) and distal (D) stumps of the sciatic nerve. Bar graphs (lower panel in (a) and (b)) represent quantification of relative band intensity to actin. Error bar (mean ± SEM, n=4). ∗∗∗ p
    Anti Il 6, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 729 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam 6 well plates
    Rabeprazole promotes topo I degradation and irinotecan resistance. A , HCT116 cells were plated in a <t>6-well</t> plate and treated with various concentrations of rabeprazole (0, 10, 20 μM) for 72 h, and then with 2.5 μM SN-38 and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin. B , Genomically edited HCT116 cells with TopoI-EGFP fusion proteins were treated with 5 and 10 μM of Rabeprazole for 48 hours and topoI-GFP protein level was analyzed by confocal microscope. C . HCT116 cells were plated in a 6-well plate, treated with 40 μM rabeprazole or DMSO for 72 h, and then with 2.5 μM SN-38, and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-pDNA-PKcs and anti-β-actin. D , HCT 116 cells were treated with rabeprazole, control and treated cells were analyzed by immunofluorescence analysis with anti-phospho-DNA-PKcs-pS2056 and confocal microscopy. E . HCT116 cells were plated in a 6-well plate and treated with rabeprazole or DMSO for 72 h. Then, 50 cells were plated in each well of a 6-well plate and treated with various concentrations of SN-38 for 24 hours. Cell colonies were counted after 14 days. F . HCT116 cells were plated in a 6-well plate and treated with 40 μM rabeprazole or DMSO for 72 h. Then, cells were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by luminescence detection.
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    93
    Thermo Fisher human il 6
    STAT3 nuclear translocation occurs independently of tyrosine phosphorylation. ( A ) HeLa cells (lanes 1 and 2) and HeLa cells transfected with STAT3–GFP (lanes 3 and 4) were untreated (-) or treated (+) with human <t>IL-6</t> for 30 min. Whole-cell lysates
    Human Il 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 580 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech recombinant human il 6
    Hierarchical clustering of autophagy-related genes in <t>IL-6-</t> and TNF-α-treated placental explants. Human first-trimester placental explants from different patients ( n = 8) were incubated with or without IL-6 (10 ng/ml) and TNF-α (10 ng/ml), respectively, for 48h. RNA from each treatment group was pooled and subjected to autophagy qPCR array. The heat map with dendrograms shows log2 fold-changes in blue for downregulated and in red for upregulated autophagy-related genes in IL-6-treated ( left column ) and TNF-α- treated ( right column ) placental explants relative to untreated controls
    Recombinant Human Il 6, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 661 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems rmil 6
    IL-6 enhances neural stem cell expansion/maintenance, which can be inhibited by COX inhibitors. Primary neurospheres were dissociated and replated into either control media or media supplemented with 5ng/mL of <t>rmIL-6.</t> The number (A) and size (B) of spheres was quantified after 6 days. IL-6 increased the number of secondary neurospheres (*, p
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    Becton Dickinson mouse il 6
    <t>IL-6</t> induction in mouse macrophage cell line J774A.1 by PagL LPS (A) and LpxL1 LPS (B). Tested were free LPS, liposomes with incorporated LPS, and liposomes with coincorporated LPS and OpaJ at LPS/protein ratios of 1:5 and 1:50 (μmol/μg).
    Mouse Il 6, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 334 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant il 6
    MSCs decrease the splenic Th17/Treg ratio and mRNA levels of pro-inflammatory cytokines in the ankle joint. (A, B) CD4, CD25, FoxP3, and RORγt expression in the spleen were determined by flow cytometry. The frequency of Th17 cells was reported as the percentage of CD4+ RORγt+ cells out of the total CD4+ T cell population. The frequency of Treg was reported as the percentage of CD4+CD25+FoxP3+ cells out of the total CD4+ T cell population. (C) Total RNA was isolated from the joints, and the expression of human, mouse IL-1Ra and pro-inflammatory cytokines such as IL-1β, <t>IL-6,</t> TNF-α, and IL-17 was investigated. (D-I) The densitometric quantification of the mRNA expression in joints. β-actin expression was used as the endogenous control. Data represent the mean ± SEM. *, P
    Recombinant Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 470 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam interleukin 6
    Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic <t>IL-6</t> levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P
    Interleukin 6, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher il 6 mouse elisa kit
    Determination of levels of TNF-α and <t>IL-6</t> in small intestines of AG129 mice after inoculation of ICs. Groups of AG129 mice ( n = 3) were challenged with a sub-lethal dose of DENV-2 S221 (V) or 100% in vitro -neutralized ICs of DENV-2 S221 with mAb 4G2 (V+4G2 ICs), anti-mE1E2 bv VLP antiserum (V+α-mE1E2 bv VLP ICs), anti-pmE1+E2 VLP antiserum (V+ α-pmE1+E2 VLP ICs), anti-mE3E4 bv VLP antiserum (V+α-mE3E4 bv VLP ICs), or anti-DENV-2 S221 antiserum (V+ α-DENV-2 S221 ICs), in parallel with the inoculations described in Figure 6B . One group was included as negative control (NC) which did not receive any treatment. Three days post-inoculation mice were euthanized, small intestines dissected out after perfusion and homogenized. Clarified homogenates were used for the determination of TNF-α (A) and IL-6 (B) using commercial <t>ELISA</t> kits, with purified recombinant murine TNF-α and IL-6 as references. Data shown are the mean values with the bars denoting standard deviation ( n = 3).
    Il 6 Mouse Elisa Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti il6 antibody
    The expression of inflammatory cytokines and fibrotic factors are reduced in STZ-induced diabetic rats. ( A ) The kidney tissues were fixed in formalin and then subjected to immunofluorescence detection of TNF-α (arrow heads pointing to dark-brown dots indicating TNF-α expression). n = 5 per group, ( B ) <t>IL6</t> protein level ( C ) MCP1 protein level ( D ) TGF-β mRNA level, ( E ) TGF-β protein level with a representative blot, ( F ) Fibronectin (FN) mRNA level in kidney tissues. Data are shown as the means ± SEM *p
    Anti Il6 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 270 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti interleukin 6
    Immunohistochemistry reactions for analysis of the expression of interleukins (IL-6 and IL-10). (A, B and C), tissues marked with primary antibody <t>anti-interleukin-6</t> (sc-130326); (D, E and F), tissues marked with primary antibody anti-interleukin-10 (H-160).
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    Bio-Rad interleukin 6
    Supplementation with  Lactobacillus reuteri  100-23 and  Lactobacillus gasseri  311476 mixture reduces inflammatory cytokines. A–F . Plasma levels of interleukin 4 (IL-4), interleukin 10 (IL-10), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8), and granulocyte colony-stimulating factor (G-CSF) in control mice (CT), in mice receiving lactobacilli (Lrg), in mice transplanted with BaF3 cells (BaF3) and in mice transplanted with BaF3 cells and receiving lactobacilli (BaF3-Lrg). Data with different superscript letters are significantly different (p
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    R&D Systems human recombinant il 6
    Arsenic induced miR-301a expression dependent on <t>IL-6/STAT3</t> signaling. (A) BEAS-2B cells were treated with arsenic (5 and 10 μM) for 24 h and with H2O2 as positive control. The levels of IL-6 in the culture medium was determined by using ELISA Kit. (B) BEAS-2B cells were treated with different concentrations of IL-6 (0, 10, 50, and 100 ng/mL) and IL-6 plus Stat3 inhibitor (S31-201, 30 μM), at 6 h and the expression of miR-301a was determined by qPCR. (C) BEAS-2B cells were treated with arsenic (As, 10 μM), arsenic (As) and S31-201, arsenic (As) and IL-6 neutralizing antibody (anti-IL-6) at 6 h and the expression of miR-301a was determined by qPCR. (D) BEAS-2B cells were transfected with Anti-control (Anti-control) and LNA-Anti-miR-301a(Anti-miR-301a). After 48 h post transfection, cells were treated with arsenic (5 μM and 10 μM) for 24 h and western blot analyses of phospho-Stat3 (pStat3), Stat3, IκBα expression. Gel pictures were acquired by using Tanon5200 and the exposure time was 60 sec for pStat3, 20 sec for Stat3, 20 sec for IκBα, and 5 sec for β-actin. (E) Protein expression from (D) were quantified by band intensity and normalized to β-actin. ( F ) pStat3 expression was quantified by band intensity and normalized to total Stat3. Values represented the mean ± SD of three independent experiments. *P
    Human Recombinant Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 272 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Meso Scale Diagnostics LLC interleukin 6
    a - f Mitochondrial proteins and inflammatory cytokines levels in explant cultured media in SIM tissue and surrounding matched-normal tissue. Wilcoxon matched-pairs signed rank tests demonstrated significantly increased levels of a cytochrome c (n=12), b SMAC/Diablo (n=8), c IL-1beta (n=12), d <t>IL-6</t> (n=12), e IL-8 (n=12) and f TNF-alpha (n=12) in SIM tissue compared to surrounding normal epithelium. * p ≤ 0.05, ** p ≤ 0.005 and *** p ≤ 0.0005
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    Image Search Results


    Secretion of interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) from BM-MSCs after stimulation with sEV-OCS or sEV-OCS-Cis. ( A – C ) Secretion levels of ( A ) IL-6, ( B ) IL-8, and ( C ) VEGFA of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis measured by multiplex fluorescent bead-based immunoassay analysis. BM-MSCs cell-cultured media (CM) was harvested 24 hours post-stimulation with sEV-OCS (CM MCS -sEV-OCS) or sEV-OCS-Cis (CM MCS -sEV-OCS-Cis). CM of BM-MSCs without sEV stimulation (CM MCS -non-sEV) was used as control. Results are mean ± SEM of n = 3. Statistical analysis was performed using the Kruskal–Wallis test followed by the Mann–Whitney test. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Small Extracellular Vesicles Released from Ovarian Cancer Spheroids in Response to Cisplatin Promote the Pro-Tumorigenic Activity of Mesenchymal Stem Cells

    doi: 10.3390/ijms20204972

    Figure Lengend Snippet: Secretion of interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) from BM-MSCs after stimulation with sEV-OCS or sEV-OCS-Cis. ( A – C ) Secretion levels of ( A ) IL-6, ( B ) IL-8, and ( C ) VEGFA of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis measured by multiplex fluorescent bead-based immunoassay analysis. BM-MSCs cell-cultured media (CM) was harvested 24 hours post-stimulation with sEV-OCS (CM MCS -sEV-OCS) or sEV-OCS-Cis (CM MCS -sEV-OCS-Cis). CM of BM-MSCs without sEV stimulation (CM MCS -non-sEV) was used as control. Results are mean ± SEM of n = 3. Statistical analysis was performed using the Kruskal–Wallis test followed by the Mann–Whitney test. * p

    Article Snippet: The concentrations of interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) were determined in CM of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis for 24 hours using a MAGPIX System (ThermoFisher Scientific, Waltham, MA, USA) as was described previously [ ].

    Techniques: Multiplex Assay, Bead-based Assay, Cell Culture, MANN-WHITNEY

    Concentrations of ( a ) IL-1β, ( b ) IL-6, ( c ) TNF-α ( d ) anti-OVA Ab in serum determined by ELISA. The results are representative of means ± SD ( n = 5; * p

    Journal: Polymers

    Article Title: Hydrogel is Superior to Fibrin Gel as Matrix of Stem Cells in Alleviating Antigen-Induced Arthritis

    doi: 10.3390/polym8050182

    Figure Lengend Snippet: Concentrations of ( a ) IL-1β, ( b ) IL-6, ( c ) TNF-α ( d ) anti-OVA Ab in serum determined by ELISA. The results are representative of means ± SD ( n = 5; * p

    Article Snippet: Percoll density gradient, thrombin, OVA and interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and anti-ovalbumin antibody (anti-OVA Ab) ELISA assay kits and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich (Shanghai, China).

    Techniques: Enzyme-linked Immunosorbent Assay

    JSI-124 inhibited bleomycin-induced bronchoalveolar inflammatory cell extravasation and lung fibrosis. Wistar rats were administered a single intratracheal dose of bleomycin (BLM; 3.75 U/kg) on day 1. JSI-124 (1 mg/kg/day) or vehicle was administered intraperitoneally from day 14 until the analysis at day 28 ( n = 10 per group). Total bronchoalveolar lavage ( a ) protein, ( b , c ) inflammatory cells, ( d ) lung weight, ( e ) protein, and ( f , g ) IL-6 and IL-13 content were measured at day 28. h Masson’s trichrome (upper panels, scale bar: 100 μm) of control, BLM, and BLM + JSI-124 tissue. i Fibrosis Ashcroft scores were assessed as described in the Methods. j Micro-CT images were acquired on day 28 and ( k ) quantified as Hounsfield units. The results are expressed as the mean ± SEM, n = 10. Statistical significance was assessed using a t -test or one-way ANOVA followed by a Bonferroni post-hoc test. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: JSI-124 inhibited bleomycin-induced bronchoalveolar inflammatory cell extravasation and lung fibrosis. Wistar rats were administered a single intratracheal dose of bleomycin (BLM; 3.75 U/kg) on day 1. JSI-124 (1 mg/kg/day) or vehicle was administered intraperitoneally from day 14 until the analysis at day 28 ( n = 10 per group). Total bronchoalveolar lavage ( a ) protein, ( b , c ) inflammatory cells, ( d ) lung weight, ( e ) protein, and ( f , g ) IL-6 and IL-13 content were measured at day 28. h Masson’s trichrome (upper panels, scale bar: 100 μm) of control, BLM, and BLM + JSI-124 tissue. i Fibrosis Ashcroft scores were assessed as described in the Methods. j Micro-CT images were acquired on day 28 and ( k ) quantified as Hounsfield units. The results are expressed as the mean ± SEM, n = 10. Statistical significance was assessed using a t -test or one-way ANOVA followed by a Bonferroni post-hoc test. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Micro-CT

    Effects of JAK2 and STAT3 on ATII to mesenchymal and fibroblast to myofibroblast transitions. Primary ATII and lung fibroblasts were isolated from the lungs of IPF patients. a The cells were incubated with the dual p-JAK2/p-STAT3 inhibitor JSI-124 for 30 min followed by TGF-β1 stimulation for an additional 24 h. IL-6 and IL-13 levels in cell supernatants were measured using ELISA. b Ratios of JAK2/p-JAK2 and STAT3/p-STAT3 were determined by western blotting in ATII cells stimulated for 40 min or 24 h with TGF-β1 in the presence or absence of JSI-124. c , d ATII cells were pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124, and then stimulated for 72 h with TGF-β1 ( c ) or IL-6/IL-13 ( d ). e Levels of IL-6 and IL-13 in primary fibroblasts. f JAK2/p-JAK2 and STAT3/p-STAT3 protein expression in human lung fibroblasts. g , h Primary lung fibroblasts pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and stimulated for 72 h with TGF-β1 ( g ) or IL-6/IL-13 ( h ). Representative western blots are shown. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: Effects of JAK2 and STAT3 on ATII to mesenchymal and fibroblast to myofibroblast transitions. Primary ATII and lung fibroblasts were isolated from the lungs of IPF patients. a The cells were incubated with the dual p-JAK2/p-STAT3 inhibitor JSI-124 for 30 min followed by TGF-β1 stimulation for an additional 24 h. IL-6 and IL-13 levels in cell supernatants were measured using ELISA. b Ratios of JAK2/p-JAK2 and STAT3/p-STAT3 were determined by western blotting in ATII cells stimulated for 40 min or 24 h with TGF-β1 in the presence or absence of JSI-124. c , d ATII cells were pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124, and then stimulated for 72 h with TGF-β1 ( c ) or IL-6/IL-13 ( d ). e Levels of IL-6 and IL-13 in primary fibroblasts. f JAK2/p-JAK2 and STAT3/p-STAT3 protein expression in human lung fibroblasts. g , h Primary lung fibroblasts pre-incubated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and stimulated for 72 h with TGF-β1 ( g ) or IL-6/IL-13 ( h ). Representative western blots are shown. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Isolation, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

    Fibroblast migration and proliferation in IPF were dependent on JAK2 and STAT3 activation. a , b Primary human fibroblasts from IPF patients were pre-treated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and then cell migration was assessed. A scrape-wound was created by using a sterile p200 pipette tip to make a perpendicular linear scratch in the culture. After the cells had been washed, culture medium with or without pharmacologic modulators and IL-6/IL-13 was added. The size of the wound remaining was analyzed at the indicated times and expressed as a percentage of the initial wound area. c Fibroblast proliferation during 48 h was evaluated by the BrdU assay. Different JAK2 and STAT3 inhibitors were added 30 min before 10% fetal bovine serum (FBS) or ( c ) 50 ng IL-6/IL-13/mL ( d ) was added as the stimulus. Values are expressed as relative absorbance (450 nm) units. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: Fibroblast migration and proliferation in IPF were dependent on JAK2 and STAT3 activation. a , b Primary human fibroblasts from IPF patients were pre-treated for 30 min with 1 μM of the p-STAT3 inhibitor 5,15 DPP, the p-JAK2 inhibitor NSC33994, or the dual p-JAK2/p-STAT3 inhibitor JSI-124 and then cell migration was assessed. A scrape-wound was created by using a sterile p200 pipette tip to make a perpendicular linear scratch in the culture. After the cells had been washed, culture medium with or without pharmacologic modulators and IL-6/IL-13 was added. The size of the wound remaining was analyzed at the indicated times and expressed as a percentage of the initial wound area. c Fibroblast proliferation during 48 h was evaluated by the BrdU assay. Different JAK2 and STAT3 inhibitors were added 30 min before 10% fetal bovine serum (FBS) or ( c ) 50 ng IL-6/IL-13/mL ( d ) was added as the stimulus. Values are expressed as relative absorbance (450 nm) units. The results are expressed as the mean (SEM) of n = 4 (cells from four IPF patients) experiments per condition. Two-way ANOVA followed by post-hoc Bonferroni tests. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Migration, Activation Assay, Transferring, BrdU Staining

    Dual JAK2/STAT3 gene silencing suppressed ATII to mesenchymal and fibroblast to myofibroblast transitions. ATII A549 and human lung fibroblast MRC5 cell lines were transfected with control siRNA(−), siRNA-JAK2, siRNA-STAT3, or both siRNA-JAK2/STAT3 and stimulated for 72 h with TG-Fβ1 or IL-6/IL-13 at a concentration of 5 ng/mL. a Total protein and RNA from cell lysates were analyzed by ( a , b ) western blot and ( c ) qPCR. Mesenchymal collagen type I and epithelial E-cadherin markers were measured using ( a ) western blot and ( c ) qPCR. Senescence p21, autophagy LC3I/II, and anti-apoptotic BCL-2 markers were ( a ) measured using western blot and ( b ) quantified by densitometry. Data are expressed as the ratio to β-actin protein and mRNA levels. The results are expressed as the mean (SEM) of n = 4 independent experiments per condition. One-way ANOVA followed by post-hoc Bonferroni tests. * P

    Journal: Respiratory Research

    Article Title: The JAK2 pathway is activated in idiopathic pulmonary fibrosis

    doi: 10.1186/s12931-018-0728-9

    Figure Lengend Snippet: Dual JAK2/STAT3 gene silencing suppressed ATII to mesenchymal and fibroblast to myofibroblast transitions. ATII A549 and human lung fibroblast MRC5 cell lines were transfected with control siRNA(−), siRNA-JAK2, siRNA-STAT3, or both siRNA-JAK2/STAT3 and stimulated for 72 h with TG-Fβ1 or IL-6/IL-13 at a concentration of 5 ng/mL. a Total protein and RNA from cell lysates were analyzed by ( a , b ) western blot and ( c ) qPCR. Mesenchymal collagen type I and epithelial E-cadherin markers were measured using ( a ) western blot and ( c ) qPCR. Senescence p21, autophagy LC3I/II, and anti-apoptotic BCL-2 markers were ( a ) measured using western blot and ( b ) quantified by densitometry. Data are expressed as the ratio to β-actin protein and mRNA levels. The results are expressed as the mean (SEM) of n = 4 independent experiments per condition. One-way ANOVA followed by post-hoc Bonferroni tests. * P

    Article Snippet: IL-6 and IL-13 cytokines were analyzed in the cell culture supernatants of human ATII and fibroblast using commercially available Quantikine® ELISA kits for human IL-6 (Catalog No. D6050; R & D Systems, Madrid, Spain) and IL-13 (Catalog No. D1300B), and in the bronchoalveolar lavage (BAL) fluid of rats using the ELISA rat IL-6 (Catalog No. KRC0061; Invitrogen™, Madrid, Spain) and IL-13 (Catalog No. KRC0132; Invitrogen™) kits according to the manufacturers’ protocols.

    Techniques: Transfection, Concentration Assay, Western Blot, Real-time Polymerase Chain Reaction

    LPS-Trap-Fc1 to -Fc4 bind LPS and block LPS-mediated IL-6 production in Mono Mac 6 cells. (A) Supernatants of HEK293T cells transfected with LPS-Trap-Fc1 were incubated in the presence (lane 1) or absence (lane 3) of biotin-LPS. Additionally, supernatants were preincubated with an anti-TLR4/MD-2 MAb (lane 2) or an excess of LPS (lane 4). Complexes were immunoprecipitated with streptavidin-Sepharose, subjected to SDS-PAGE, and analyzed by Western blotting with an anti-FLAG MAb. (B) RAW 264.7 cells were incubated with LPS-Trap-Fc1 or a medium control. Cells were stimulated with 100 ng/ml LPS overnight, and IL-6 levels in supernatants were determined by ELISA. *, P

    Journal: Infection and Immunity

    Article Title: Lipopolysaccharide-Trap-Fc, a Multifunctional Agent To Battle Gram-Negative Bacteria ▿

    doi: 10.1128/IAI.00004-09

    Figure Lengend Snippet: LPS-Trap-Fc1 to -Fc4 bind LPS and block LPS-mediated IL-6 production in Mono Mac 6 cells. (A) Supernatants of HEK293T cells transfected with LPS-Trap-Fc1 were incubated in the presence (lane 1) or absence (lane 3) of biotin-LPS. Additionally, supernatants were preincubated with an anti-TLR4/MD-2 MAb (lane 2) or an excess of LPS (lane 4). Complexes were immunoprecipitated with streptavidin-Sepharose, subjected to SDS-PAGE, and analyzed by Western blotting with an anti-FLAG MAb. (B) RAW 264.7 cells were incubated with LPS-Trap-Fc1 or a medium control. Cells were stimulated with 100 ng/ml LPS overnight, and IL-6 levels in supernatants were determined by ELISA. *, P

    Article Snippet: The supernatants were tested for secreted interleukin-6 (IL-6) by ELISA (Becton Dickinson, Heidelberg, Germany).

    Techniques: Blocking Assay, Transfection, Incubation, Immunoprecipitation, SDS Page, Western Blot, Enzyme-linked Immunosorbent Assay

    Influence of erythropoietin on meningococcal-induced cytokine production. Interleukin (IL)-6 and tumour necrosis factor (TNF)-α-positive monocytes are shown in neonates ( n = 20), infants (defined as

    Journal:

    Article Title: Attenuation of monocyte proinflammatory cytokine responses to Neisseria meningitidis in children by erythropoietin

    doi: 10.1111/j.1365-2249.2008.03760.x

    Figure Lengend Snippet: Influence of erythropoietin on meningococcal-induced cytokine production. Interleukin (IL)-6 and tumour necrosis factor (TNF)-α-positive monocytes are shown in neonates ( n = 20), infants (defined as

    Article Snippet: Because non-haematopoietic effects of erythropoietin require higher concentrations [ ] we employed human recombinant erythropoietin beta (NeoRecormon, Firma Roche, Grenzen-Wylach, Germany), diluted in RPMI-1640 in final concentrations of 100 and 500 U/ml respectively, which was added to the cultures 1 h prior to stimulation with heat-inactivated meningococcal strains at a final concentration of 1·2 × 108 per ml for 5 h. As a positive control, lipopolysaccharide (LPS) obtained from Escherichia coli ( E. coli 0127: B8; no. L3129; Sigma, Deisenhofen, Germany) was added at a concentration of 30 ng/ml for 5 h. Cells were washed in Hanks's balanced salt solution (HBSS) and resuspended in a buffer consisting of HBSS, 0·1% saponin (Riedel de Haen, Hanover, Germany) and 0·01 M HEPES buffer (Seromed Biochrome, Berlin, Germany); 200 µl aliquots of cells were added to tubes containing 0·5 µg/10 µl of monoclonal antibodies (mAbs) against CD14, tumour necrosis factor (TNF)-α and interleukin (IL)-6 (Becton Dickinson, Heidelberg, Germany).

    Techniques:

    IL-6 is up-regulated in PDGFRα + stromal cells after SO 2 injury. ( A ) RNAs were extracted from whole trachea at 0, 1, 2, and 14 d after injury and subjected to quantitative RT-PCR analysis. The mRNA expression level of cytokines was normalized to Gapdh . ( B ) In situ hybridization combined with immunohistochemistry shows that Il-6 mRNA (red) is expressed in cells in the stroma beneath basal cells (K5 + , green) after SO 2 injury. ( C ) Quantitative PCR analysis of Il-6 expression in sorted stromal cells [ Pdgfr α ( Pdgfra )-GFP + ] and immune cell subpopulations from the trachea at 24 hpi. ( D ) Immunohistochemistry of a trachea section at 24 hpi shows Pdgfra -GFP + cells (GFP + , green) in the stroma beneath the epithelium with basal cells (K5 + , red). ( E ) In situ hybridization and immunohistochemistry show that Pdgfra -GFP + cells (GFP + , green) express Il-6 mRNA (red) at 24 hpi. (Scale bars: B and E , 20 μm; D , 50 μm.) * P

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

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6 is up-regulated in PDGFRα + stromal cells after SO 2 injury. ( A ) RNAs were extracted from whole trachea at 0, 1, 2, and 14 d after injury and subjected to quantitative RT-PCR analysis. The mRNA expression level of cytokines was normalized to Gapdh . ( B ) In situ hybridization combined with immunohistochemistry shows that Il-6 mRNA (red) is expressed in cells in the stroma beneath basal cells (K5 + , green) after SO 2 injury. ( C ) Quantitative PCR analysis of Il-6 expression in sorted stromal cells [ Pdgfr α ( Pdgfra )-GFP + ] and immune cell subpopulations from the trachea at 24 hpi. ( D ) Immunohistochemistry of a trachea section at 24 hpi shows Pdgfra -GFP + cells (GFP + , green) in the stroma beneath the epithelium with basal cells (K5 + , red). ( E ) In situ hybridization and immunohistochemistry show that Pdgfra -GFP + cells (GFP + , green) express Il-6 mRNA (red) at 24 hpi. (Scale bars: B and E , 20 μm; D , 50 μm.) * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Quantitative RT-PCR, Expressing, In Situ Hybridization, Immunohistochemistry, Real-time Polymerase Chain Reaction

    IL-6/STAT3 signaling is activated in tracheal epithelium during repair. ( A ) Schematic of the SO 2 injury model. After exposure to SO 2 , luminal cells die. Basal cells spread, proliferate, and generate early progenitors. These progenitors differentiate into ciliated and secretory cells, and repair is complete in 2 wk. ( B ) Longitudinal midline sections stained with antibodies to p-STAT3 (red) and p63 (green), a marker for basal cells. ( C ) Expression of p-STAT3 (red) and FOXJ1 (green) during epithelial repair. Note the coexpression of p-STAT3 and FOXJ1 at 3 dpi. (Scale bars: B and C .)

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

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6/STAT3 signaling is activated in tracheal epithelium during repair. ( A ) Schematic of the SO 2 injury model. After exposure to SO 2 , luminal cells die. Basal cells spread, proliferate, and generate early progenitors. These progenitors differentiate into ciliated and secretory cells, and repair is complete in 2 wk. ( B ) Longitudinal midline sections stained with antibodies to p-STAT3 (red) and p63 (green), a marker for basal cells. ( C ) Expression of p-STAT3 (red) and FOXJ1 (green) during epithelial repair. Note the coexpression of p-STAT3 and FOXJ1 at 3 dpi. (Scale bars: B and C .)

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Staining, Marker, Expressing

    Effect of IL-6/STAT3 on tracheal epithelial repair in vivo. ( A ) Schematic of gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) model. Floxed alleles are deleted, and the YFP reporter is activated in basal cells with three doses of Tmx. One week later, mice are exposed to SO 2 and tracheas are harvested at 6 dpi. ( B ) Representative midline sections of tracheas (ventral) stained with YFP (lineage label, green) and a-tub (ciliated cells, red) in control ( K5-CreER T2 ; Rosa-YFP ) and gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) mice. A similar analysis was carried out using antibodies to K5 for basal cells and SCGB1A1 and SCGB3A2 for secretory cells, respectively. ( C ) Percentage of total lineage-labeled cells (YFP + ) throughout the trachea that are ciliated, secretory, or basal cells. Blue and red bars show K5-CreER T2 ; Rosa-YFP and K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP , respectively. ( D ) FOXJ1 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( E ) SCGB3A2 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( F ) In Il-6 null mice, there is a reduction of ciliated cells (FOXJ1 + ) and an increase of secretory cells (SCGB3A2 + ) after SO 2 injury (4 dpi). * P

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

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: Effect of IL-6/STAT3 on tracheal epithelial repair in vivo. ( A ) Schematic of gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) model. Floxed alleles are deleted, and the YFP reporter is activated in basal cells with three doses of Tmx. One week later, mice are exposed to SO 2 and tracheas are harvested at 6 dpi. ( B ) Representative midline sections of tracheas (ventral) stained with YFP (lineage label, green) and a-tub (ciliated cells, red) in control ( K5-CreER T2 ; Rosa-YFP ) and gain-of-function ( K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP ) mice. A similar analysis was carried out using antibodies to K5 for basal cells and SCGB1A1 and SCGB3A2 for secretory cells, respectively. ( C ) Percentage of total lineage-labeled cells (YFP + ) throughout the trachea that are ciliated, secretory, or basal cells. Blue and red bars show K5-CreER T2 ; Rosa-YFP and K5-CreER T2 ; Socs3 flox/flox ; Rosa-YFP , respectively. ( D ) FOXJ1 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( E ) SCGB3A2 staining (green) of airway epithelium at 4 dpi in WT and Il-6 null mice. ( F ) In Il-6 null mice, there is a reduction of ciliated cells (FOXJ1 + ) and an increase of secretory cells (SCGB3A2 + ) after SO 2 injury (4 dpi). * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: In Vivo, Mouse Assay, Staining, Labeling

    Effect of IL-6 on regeneration of human epithelium in ALI culture. ( A ) Schematic of ALI culture of primary HBE cells. ( B ) Whole-mount staining of day 21 cultures for ciliated (α-tubulin, green) and secretory (SCGB3A1, red) cells. Nuclei are blue (DAPI). (Scale bar: 100 μm.) ( C ) Quantification of whole-mount staining, shown as a fold change over untreated culture. The α-tubulin + or SCGB3A1 + cells were counted in four randomly chosen areas (0.18 mm 2 ) per filter. Values are mean ± SD for cultures from three different donors. * P

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

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: Effect of IL-6 on regeneration of human epithelium in ALI culture. ( A ) Schematic of ALI culture of primary HBE cells. ( B ) Whole-mount staining of day 21 cultures for ciliated (α-tubulin, green) and secretory (SCGB3A1, red) cells. Nuclei are blue (DAPI). (Scale bar: 100 μm.) ( C ) Quantification of whole-mount staining, shown as a fold change over untreated culture. The α-tubulin + or SCGB3A1 + cells were counted in four randomly chosen areas (0.18 mm 2 ) per filter. Values are mean ± SD for cultures from three different donors. * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Staining

    IL-6 enhances Foxj1-GFP expression in the mouse tracheosphere culture assay. ( A ) Schematic of the assay. NGFR + basal cells from Foxj1-GFP tracheas were cultured in 50% Matrigel in 96-well inserts. ( Right ) Section of a typical sphere with acetylated tubulin + (a-tub) ciliated (magenta) and Splunc + secretory cells (green). IHC, immunohistochemistry. The effect of IL-6 ( B ) and STAT3 inhibitor ( C ) on Foxj1 -GFP expression is shown. Differential interference contrast images ( Upper ) and fluorescent images ( Lower ) of the same spheres are shown. ( D ) Quantification by FACS at day 11 of the percentage of GFP + cells in dissociated spheres treated with IL-6 (0, 1, and 10 ng/mL). ( E ) Quantification at different times of GFP + cells in spheres cultured with or without IL-6 (1 ng/mL). ( F ) Representative sections of spheres at day 14 treated with IL-6 ( Left , 10 ng/mL) or S3I-201 ( Right , 200 μM, days 4–7). Both sections were stained with antibodies to a-tub + (magenta) and Splunc + (green). * P

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

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6 enhances Foxj1-GFP expression in the mouse tracheosphere culture assay. ( A ) Schematic of the assay. NGFR + basal cells from Foxj1-GFP tracheas were cultured in 50% Matrigel in 96-well inserts. ( Right ) Section of a typical sphere with acetylated tubulin + (a-tub) ciliated (magenta) and Splunc + secretory cells (green). IHC, immunohistochemistry. The effect of IL-6 ( B ) and STAT3 inhibitor ( C ) on Foxj1 -GFP expression is shown. Differential interference contrast images ( Upper ) and fluorescent images ( Lower ) of the same spheres are shown. ( D ) Quantification by FACS at day 11 of the percentage of GFP + cells in dissociated spheres treated with IL-6 (0, 1, and 10 ng/mL). ( E ) Quantification at different times of GFP + cells in spheres cultured with or without IL-6 (1 ng/mL). ( F ) Representative sections of spheres at day 14 treated with IL-6 ( Left , 10 ng/mL) or S3I-201 ( Right , 200 μM, days 4–7). Both sections were stained with antibodies to a-tub + (magenta) and Splunc + (green). * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Expressing, Cell Culture, Immunohistochemistry, FACS, Staining

    IL-6 enhances expression of cilia-related genes and inhibits Notch1 expression in mouse ALI culture. ( A ) Schematic of ALI culture of mouse tracheal epithelial cells. At day 7, IL-6 (10 ng/mL) was added to culture medium in the lower chamber. Cells were harvested after 6, 12, and 24 h, and total RNA was extracted. ( B ) Quantitative RT-PCR shows that IL-6 treatment promotes the expression of the known target gene Socs3 and ciliogenesis-related genes, such as Multicilin ( Mcidas) and Foxj1 . IL-6 treatment also inhibits Notch1 and promotes expression of Cdc20b , the host gene for miR-449a/b. No significant changes were observed in the expression of Notch2 , Dll1 , or Jagged1 . ( C ) ChIP assay shows that p-STAT3 binding to promoter regions of Socs3, Foxj1 , Mcidas , and Notch1 is increased after IL-6 stimulation. * P

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

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: IL-6 enhances expression of cilia-related genes and inhibits Notch1 expression in mouse ALI culture. ( A ) Schematic of ALI culture of mouse tracheal epithelial cells. At day 7, IL-6 (10 ng/mL) was added to culture medium in the lower chamber. Cells were harvested after 6, 12, and 24 h, and total RNA was extracted. ( B ) Quantitative RT-PCR shows that IL-6 treatment promotes the expression of the known target gene Socs3 and ciliogenesis-related genes, such as Multicilin ( Mcidas) and Foxj1 . IL-6 treatment also inhibits Notch1 and promotes expression of Cdc20b , the host gene for miR-449a/b. No significant changes were observed in the expression of Notch2 , Dll1 , or Jagged1 . ( C ) ChIP assay shows that p-STAT3 binding to promoter regions of Socs3, Foxj1 , Mcidas , and Notch1 is increased after IL-6 stimulation. * P

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques: Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation, Binding Assay

    Model for regulation of ciliogenesis in airway epithelium by STAT3. ( Upper ) After injury, STAT3 in both basal cells and progenitors is activated by IL-6 secreted from PDGFRα + stromal cells. Ciliogenesis is likely promoted both at the level of cell fate determination and at the level of differentiation/maturation of the progenitors of multiciliated cells. ( Lower ) Schematic model for how STAT3 may directly regulate ciliogenesis-related genes during repair of the tracheal epithelium.

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

    Article Title: IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells

    doi: 10.1073/pnas.1409781111

    Figure Lengend Snippet: Model for regulation of ciliogenesis in airway epithelium by STAT3. ( Upper ) After injury, STAT3 in both basal cells and progenitors is activated by IL-6 secreted from PDGFRα + stromal cells. Ciliogenesis is likely promoted both at the level of cell fate determination and at the level of differentiation/maturation of the progenitors of multiciliated cells. ( Lower ) Schematic model for how STAT3 may directly regulate ciliogenesis-related genes during repair of the tracheal epithelium.

    Article Snippet: Mouse ALI cultures at day 7 were exposed to mouse IL-6 (20 ng/mL; R & D Systems) at 37 °C for 4 h. Approximately 4 × 106 cells were fixed at room temperature for 10 min and scraped off the inserts.

    Techniques:

    Niphatenone B enantiomers block transactivation of AR NTD. (A) Chemical structures of sintokamide, EPI-001, and niphatenone B enantiomers. (B) Transactivation assay of the AR NTD performed in LNCaP cells that were co-transfected with Gal4UAS-TATA-luciferase and AR-(1-558)-Gal4 DBD prior pre-treatment for 1 hour with 25 µM EPI-002, 7.0 µM niphatenones or DMSO vehicle control. Transactivation of AR NTD was induced by incubation with IL-6 (50 ng/ml) for 24 hours. Luciferase activity was normalized to protein concentration. (C) Niphatenones inhibit the constitutively active AR V567es splice variant. Cos-1 cells co-transfected with PB-luciferase and an expression vector for ARvar567 were treated with each niphatenone B (1 µM) or DMSO vehicle control. After 24 hours of exposure, cells were harvested and luciferase activities were normalized to protein concentrations of the samples. Data represent the mean ± SEM of n = 3 separate experiments with triplicate wells. Student's t test: ** P

    Journal: PLoS ONE

    Article Title: Characterization of Niphatenones that Inhibit Androgen Receptor N-Terminal Domain

    doi: 10.1371/journal.pone.0107991

    Figure Lengend Snippet: Niphatenone B enantiomers block transactivation of AR NTD. (A) Chemical structures of sintokamide, EPI-001, and niphatenone B enantiomers. (B) Transactivation assay of the AR NTD performed in LNCaP cells that were co-transfected with Gal4UAS-TATA-luciferase and AR-(1-558)-Gal4 DBD prior pre-treatment for 1 hour with 25 µM EPI-002, 7.0 µM niphatenones or DMSO vehicle control. Transactivation of AR NTD was induced by incubation with IL-6 (50 ng/ml) for 24 hours. Luciferase activity was normalized to protein concentration. (C) Niphatenones inhibit the constitutively active AR V567es splice variant. Cos-1 cells co-transfected with PB-luciferase and an expression vector for ARvar567 were treated with each niphatenone B (1 µM) or DMSO vehicle control. After 24 hours of exposure, cells were harvested and luciferase activities were normalized to protein concentrations of the samples. Data represent the mean ± SEM of n = 3 separate experiments with triplicate wells. Student's t test: ** P

    Article Snippet: Interleukin-6 was purchased from R & D Systems (Minneapolis, MN).

    Techniques: Blocking Assay, Transactivation Assay, Transfection, Luciferase, Incubation, Activity Assay, Protein Concentration, Variant Assay, Expressing, Plasmid Preparation

    MCR expressions during erythroblast differentiation. ( A ) Schematic diagram of an in vitro differentiation protocol for deriving erythroblasts from human HPCs. Human CD34 + HPCs were expanded for 7 days (E0–E7) and stocks were frozen in liquid nitrogen (LN 2 ). The stock cells were differentiated for 3 days (D0–D3) before undergoing maturation (M0–M7). The number of enucleated erythrocytes increased from M5 to M7. ACTH, adrenocorticotropic hormone; EPO, erythropoietin; FL, flt-3 ligand; IL-3, interleukin-3; IL-6, interleukin-6; α-MSH, α-melanocyte stimulating hormone; SCF, stem cell factor; TPO, thrombopoietin. ( B ) May-Grunwald-Giemsa staining of control erythroblasts at day M0–5 day. Cells differentiated to Pro-E stage at day M0 and Baso-E stage at day M3. Arrows, Poly-E; Arrow heads, Orho-E. *, Reticulocyte. Bar 10 μm. ( C ) Conventional RT-PCR for MCRs during erythroblasts differentiation between M0 and M6 day.

    Journal: PLoS ONE

    Article Title: Melanocortins Contribute to Sequential Differentiation and Enucleation of Human Erythroblasts via Melanocortin Receptors 1, 2 and 5

    doi: 10.1371/journal.pone.0123232

    Figure Lengend Snippet: MCR expressions during erythroblast differentiation. ( A ) Schematic diagram of an in vitro differentiation protocol for deriving erythroblasts from human HPCs. Human CD34 + HPCs were expanded for 7 days (E0–E7) and stocks were frozen in liquid nitrogen (LN 2 ). The stock cells were differentiated for 3 days (D0–D3) before undergoing maturation (M0–M7). The number of enucleated erythrocytes increased from M5 to M7. ACTH, adrenocorticotropic hormone; EPO, erythropoietin; FL, flt-3 ligand; IL-3, interleukin-3; IL-6, interleukin-6; α-MSH, α-melanocyte stimulating hormone; SCF, stem cell factor; TPO, thrombopoietin. ( B ) May-Grunwald-Giemsa staining of control erythroblasts at day M0–5 day. Cells differentiated to Pro-E stage at day M0 and Baso-E stage at day M3. Arrows, Poly-E; Arrow heads, Orho-E. *, Reticulocyte. Bar 10 μm. ( C ) Conventional RT-PCR for MCRs during erythroblasts differentiation between M0 and M6 day.

    Article Snippet: In the second passage (for differentiation; D0–D3 in ), cell stocks were thawed and cultured at 2 × 105 cells/ml in HPGM supplemented with 3 U/ml human EPO (Kyowa Hakko Kirin), 25 ng/ml SCF, 10 ng/ml recombinant human IL-3 (PeproTech), and 10 ng/ml recombinant human IL-6 (R & D Systems) for 3 days ( ).

    Techniques: In Vitro, Staining, Reverse Transcription Polymerase Chain Reaction

    Biochemical and hematological parameters of DSS-induced colitis: the colonic levels of a MPO activity, b tGSH, c IL-6 and d TNF-α; the serum levels of e IL-6 and f TNF-α; the hematologic results of (g) white blood cells. Each point represents a single sample. Asterisks and plus signs represent significant differences (*** P

    Journal: Journal of Nanobiotechnology

    Article Title: Orally administered gold nanoparticles protect against colitis by attenuating Toll-like receptor 4- and reactive oxygen/nitrogen species-mediated inflammatory responses but could induce gut dysbiosis in mice

    doi: 10.1186/s12951-018-0415-5

    Figure Lengend Snippet: Biochemical and hematological parameters of DSS-induced colitis: the colonic levels of a MPO activity, b tGSH, c IL-6 and d TNF-α; the serum levels of e IL-6 and f TNF-α; the hematologic results of (g) white blood cells. Each point represents a single sample. Asterisks and plus signs represent significant differences (*** P

    Article Snippet: The Pierce BCA protein assay kit, 2′,7′-DCF diacetate, the enhanced chemiluminescence (ECL) detection kit, and sandwich enzyme-linked immunosorbent assay (ELISA) kits for mouse IL-6 and TNF-α, DMEM, Dulbecco’s phosphate-buffered saline (DPBS) and Hank’s balanced salt solution (HBSS) were obtained from ThermoFisher Scientific (CA, USA).

    Techniques: Activity Assay

    WT-CCN1, but not DM-CCN1, induces IL-6 in macrophages and fibroblasts (A) IL-6 mRNA was measured by qRT-PCR and (B) IL-6 protein from conditioned media was measured by ELISA in serum-starved (overnight) I13.35 macrophages treated with 5 μg/ml of purified WT-CCN1 or DM-CCN1, or BSA for 24 hrs. (C) I13.35 macrophages were incubated with blocking mAbs against integrin α M (50 μg/ml) or β 2 (50 μg/ml) 1 hr prior to treatment with WT-CCN1 for 24 hrs. IL-6 mRNA was measured by qRT-PCR. (D) IL-6 mRNA was measured by qRT-PCR and (E) IL-6 protein from conditioned media was measured by ELISA in serum-starved 18Co fibroblasts treated with 5 μg/ml of purified WT-CCN1, DM-CCN1, BSA, or 25 ng/ml of TNFα for 24 hrs. (F) 18Co fibroblasts were incubated with blocking mAb against integrin α 6 for 1 hr, then treated with WT-CCN1, TNFα, or BSA for 24 hrs as above. IL-6 mRNA was measured by qRT-PCR. Data shown as mean ± SD of triplicate experiments.

    Journal: Mucosal immunology

    Article Title: The Matricellular Protein CCN1 Promotes Mucosal Healing in Murine Colitis through IL-6

    doi: 10.1038/mi.2015.19

    Figure Lengend Snippet: WT-CCN1, but not DM-CCN1, induces IL-6 in macrophages and fibroblasts (A) IL-6 mRNA was measured by qRT-PCR and (B) IL-6 protein from conditioned media was measured by ELISA in serum-starved (overnight) I13.35 macrophages treated with 5 μg/ml of purified WT-CCN1 or DM-CCN1, or BSA for 24 hrs. (C) I13.35 macrophages were incubated with blocking mAbs against integrin α M (50 μg/ml) or β 2 (50 μg/ml) 1 hr prior to treatment with WT-CCN1 for 24 hrs. IL-6 mRNA was measured by qRT-PCR. (D) IL-6 mRNA was measured by qRT-PCR and (E) IL-6 protein from conditioned media was measured by ELISA in serum-starved 18Co fibroblasts treated with 5 μg/ml of purified WT-CCN1, DM-CCN1, BSA, or 25 ng/ml of TNFα for 24 hrs. (F) 18Co fibroblasts were incubated with blocking mAb against integrin α 6 for 1 hr, then treated with WT-CCN1, TNFα, or BSA for 24 hrs as above. IL-6 mRNA was measured by qRT-PCR. Data shown as mean ± SD of triplicate experiments.

    Article Snippet: Recombinant mouse IL-6 was from Peprotech (Rocky Hill, NJ) and recombinant TNFα was from Apotech (Epalinges, Switzerland), and epidermal growth factor (EGF) from Sigma-Aldrich.

    Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Purification, Incubation, Blocking Assay

    WT-CCN1, but not DM-CCN1, induces IL-6 in vivo (A) IL-6 mRNA in the colon of wild type and Ccn1 dm/dm mice treated with 3.5% DSS for 5 days was measured by qRT-PCR, and normalized to healthy wild type colon with cyclophilin E as internal reference. Data shown as mean ± SD; n =3–4; n.s., not significant. ** P

    Journal: Mucosal immunology

    Article Title: The Matricellular Protein CCN1 Promotes Mucosal Healing in Murine Colitis through IL-6

    doi: 10.1038/mi.2015.19

    Figure Lengend Snippet: WT-CCN1, but not DM-CCN1, induces IL-6 in vivo (A) IL-6 mRNA in the colon of wild type and Ccn1 dm/dm mice treated with 3.5% DSS for 5 days was measured by qRT-PCR, and normalized to healthy wild type colon with cyclophilin E as internal reference. Data shown as mean ± SD; n =3–4; n.s., not significant. ** P

    Article Snippet: Recombinant mouse IL-6 was from Peprotech (Rocky Hill, NJ) and recombinant TNFα was from Apotech (Epalinges, Switzerland), and epidermal growth factor (EGF) from Sigma-Aldrich.

    Techniques: In Vivo, Mouse Assay, Quantitative RT-PCR

    Delivery of IL-6 enhances survival and mucosal repair of Ccn1 dm/dm mice by restoring IEC proliferation (A) Ccn1 dm/dm mice ( n =20) were injected i.p. with rIL-6 (100 ng) or vehicle control daily for 5 consecutive days after each cycle of 5% DSS feeding and were monitored for survival. ** P

    Journal: Mucosal immunology

    Article Title: The Matricellular Protein CCN1 Promotes Mucosal Healing in Murine Colitis through IL-6

    doi: 10.1038/mi.2015.19

    Figure Lengend Snippet: Delivery of IL-6 enhances survival and mucosal repair of Ccn1 dm/dm mice by restoring IEC proliferation (A) Ccn1 dm/dm mice ( n =20) were injected i.p. with rIL-6 (100 ng) or vehicle control daily for 5 consecutive days after each cycle of 5% DSS feeding and were monitored for survival. ** P

    Article Snippet: Recombinant mouse IL-6 was from Peprotech (Rocky Hill, NJ) and recombinant TNFα was from Apotech (Epalinges, Switzerland), and epidermal growth factor (EGF) from Sigma-Aldrich.

    Techniques: Mouse Assay, Injection

    Comparison of TNF- α and IL-6 production in the sciatic nerve after CCI and BGJTD/BeD treatments. Western blot analysis of TNF- α (a) and IL-6 (b) in the proximal (P) and distal (D) stumps of the sciatic nerve. Bar graphs (lower panel in (a) and (b)) represent quantification of relative band intensity to actin. Error bar (mean ± SEM, n=4). ∗∗∗ p

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Bogijetong Decoction and Its Selected Formulation Are Involved in Alleviating Neuropathic Pain in a Rat Model of Chronic Constrictive Injury

    doi: 10.1155/2018/2050636

    Figure Lengend Snippet: Comparison of TNF- α and IL-6 production in the sciatic nerve after CCI and BGJTD/BeD treatments. Western blot analysis of TNF- α (a) and IL-6 (b) in the proximal (P) and distal (D) stumps of the sciatic nerve. Bar graphs (lower panel in (a) and (b)) represent quantification of relative band intensity to actin. Error bar (mean ± SEM, n=4). ∗∗∗ p

    Article Snippet: Primary antibodies used in the present study were anti-BDNF (1:1000, mouse, monoclonal, Abcam, Cambridge, MA, USA), anti-TrkB (1:800, Rabbit, polyclonal, Abcam), anti-GAP-43 (1:800, Rabbit, polyclonal, Abcam), anti-phopsho-Erk1/2 (1:1000, Rabbit, polyclonal, Cell Signaling, Danvers, MA), anti-IL-6 (1:800, mouse, monoclonal, Abcam), anti-TNF- α (1:800, Rabbit, polyclonal, Abcam), and anti-Actin (1:20000, mouse, Sigma) antibodies, and secondary antibodies were goat anti-mouse and goat anti-rabbit horseradish peroxidase- (HRP-) conjugated antibodies (1:1000, Cell Signaling).

    Techniques: Western Blot

    Schematics of proposed mechanisms underlying BGJTD- or BeD-modulated pain responses. According to our data, signaling of TNF- α and TrkB appeared to increase after nerve ligation (a). TNF- α , which were decreased by BGJTD treatment, may cause weakening in retrograde signaling of TNFR into the cell body, leading to alleviating pain response (b). After BeD treatment, p-Erk1/2 and IL-6 signals, induced from Schwann cells and macrophages, may transmit the signals of IL-6R into the neuronal cell body (soma), inducing JAK/STAT activation and regulation of target gene expression including the repression of TrkB gene expression (c). Consequently, retrograde TrkB signaling would be attenuated and lead to alleviate pain response.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Bogijetong Decoction and Its Selected Formulation Are Involved in Alleviating Neuropathic Pain in a Rat Model of Chronic Constrictive Injury

    doi: 10.1155/2018/2050636

    Figure Lengend Snippet: Schematics of proposed mechanisms underlying BGJTD- or BeD-modulated pain responses. According to our data, signaling of TNF- α and TrkB appeared to increase after nerve ligation (a). TNF- α , which were decreased by BGJTD treatment, may cause weakening in retrograde signaling of TNFR into the cell body, leading to alleviating pain response (b). After BeD treatment, p-Erk1/2 and IL-6 signals, induced from Schwann cells and macrophages, may transmit the signals of IL-6R into the neuronal cell body (soma), inducing JAK/STAT activation and regulation of target gene expression including the repression of TrkB gene expression (c). Consequently, retrograde TrkB signaling would be attenuated and lead to alleviate pain response.

    Article Snippet: Primary antibodies used in the present study were anti-BDNF (1:1000, mouse, monoclonal, Abcam, Cambridge, MA, USA), anti-TrkB (1:800, Rabbit, polyclonal, Abcam), anti-GAP-43 (1:800, Rabbit, polyclonal, Abcam), anti-phopsho-Erk1/2 (1:1000, Rabbit, polyclonal, Cell Signaling, Danvers, MA), anti-IL-6 (1:800, mouse, monoclonal, Abcam), anti-TNF- α (1:800, Rabbit, polyclonal, Abcam), and anti-Actin (1:20000, mouse, Sigma) antibodies, and secondary antibodies were goat anti-mouse and goat anti-rabbit horseradish peroxidase- (HRP-) conjugated antibodies (1:1000, Cell Signaling).

    Techniques: Ligation, Activation Assay, Expressing

    Rabeprazole promotes topo I degradation and irinotecan resistance. A , HCT116 cells were plated in a 6-well plate and treated with various concentrations of rabeprazole (0, 10, 20 μM) for 72 h, and then with 2.5 μM SN-38 and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin. B , Genomically edited HCT116 cells with TopoI-EGFP fusion proteins were treated with 5 and 10 μM of Rabeprazole for 48 hours and topoI-GFP protein level was analyzed by confocal microscope. C . HCT116 cells were plated in a 6-well plate, treated with 40 μM rabeprazole or DMSO for 72 h, and then with 2.5 μM SN-38, and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-pDNA-PKcs and anti-β-actin. D , HCT 116 cells were treated with rabeprazole, control and treated cells were analyzed by immunofluorescence analysis with anti-phospho-DNA-PKcs-pS2056 and confocal microscopy. E . HCT116 cells were plated in a 6-well plate and treated with rabeprazole or DMSO for 72 h. Then, 50 cells were plated in each well of a 6-well plate and treated with various concentrations of SN-38 for 24 hours. Cell colonies were counted after 14 days. F . HCT116 cells were plated in a 6-well plate and treated with 40 μM rabeprazole or DMSO for 72 h. Then, cells were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by luminescence detection.

    Journal: PLoS ONE

    Article Title: CTDSP1 inhibitor rabeprazole regulates DNA-PKcs dependent topoisomerase I degradation and irinotecan drug resistance in colorectal cancer

    doi: 10.1371/journal.pone.0228002

    Figure Lengend Snippet: Rabeprazole promotes topo I degradation and irinotecan resistance. A , HCT116 cells were plated in a 6-well plate and treated with various concentrations of rabeprazole (0, 10, 20 μM) for 72 h, and then with 2.5 μM SN-38 and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin. B , Genomically edited HCT116 cells with TopoI-EGFP fusion proteins were treated with 5 and 10 μM of Rabeprazole for 48 hours and topoI-GFP protein level was analyzed by confocal microscope. C . HCT116 cells were plated in a 6-well plate, treated with 40 μM rabeprazole or DMSO for 72 h, and then with 2.5 μM SN-38, and harvested after 90 or 180 min. Cell lysates were immunoblotted with anti-pDNA-PKcs and anti-β-actin. D , HCT 116 cells were treated with rabeprazole, control and treated cells were analyzed by immunofluorescence analysis with anti-phospho-DNA-PKcs-pS2056 and confocal microscopy. E . HCT116 cells were plated in a 6-well plate and treated with rabeprazole or DMSO for 72 h. Then, 50 cells were plated in each well of a 6-well plate and treated with various concentrations of SN-38 for 24 hours. Cell colonies were counted after 14 days. F . HCT116 cells were plated in a 6-well plate and treated with 40 μM rabeprazole or DMSO for 72 h. Then, cells were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by luminescence detection.

    Article Snippet: ImmunoblottingCells cultured in 6-well plates were scraped into an ice-cold RIPA buffer.

    Techniques: Microscopy, Immunofluorescence, Confocal Microscopy

    Silencing of CTDSP1 enhances topoI degradation and irinotecan resistance. A , Cells transfected with CTDSP1 or control siRNA were lyzed and the cells’ lysates were immunoblotted with anti-CTDSP1 and anti-β-actin antibodies. B , HCT116-siRNA CTDSP1 or control siRNA, treated with 2.5 μM SN-38 were harvested after 90 and 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. Cells’ lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. C , EGFP was integrated with topoI in HCT116 cells using CRISPR/Cas9 system and CTDSP1 was knocked down in this cell line by siRNA. Cells were treated with 2.5uM SN-38 for 60 min and the topoI-EGFP signal was imaged by Leica SP5 confocal microscope. D , HCT116-siRNA-CTDSP1 or control siRNA were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by detecting the luminescence. E , HCT116-siRNA-CTDSP1 or control siRNA cells were plated in a 6-well plate and treated with various concentrations of SN-38 for 24 h. Then, 50 cells per well were plated in a 6-well plate. After 14 days, when colonies were apparent (right panel), colonies were counted and the relative number of colonies was determined (left panel).

    Journal: PLoS ONE

    Article Title: CTDSP1 inhibitor rabeprazole regulates DNA-PKcs dependent topoisomerase I degradation and irinotecan drug resistance in colorectal cancer

    doi: 10.1371/journal.pone.0228002

    Figure Lengend Snippet: Silencing of CTDSP1 enhances topoI degradation and irinotecan resistance. A , Cells transfected with CTDSP1 or control siRNA were lyzed and the cells’ lysates were immunoblotted with anti-CTDSP1 and anti-β-actin antibodies. B , HCT116-siRNA CTDSP1 or control siRNA, treated with 2.5 μM SN-38 were harvested after 90 and 180 min. Cell lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. Cells’ lysates were immunoblotted with anti-topoI and anti-β-actin antibodies. C , EGFP was integrated with topoI in HCT116 cells using CRISPR/Cas9 system and CTDSP1 was knocked down in this cell line by siRNA. Cells were treated with 2.5uM SN-38 for 60 min and the topoI-EGFP signal was imaged by Leica SP5 confocal microscope. D , HCT116-siRNA-CTDSP1 or control siRNA were plated in a 96-well plate and treated with various concentrations of SN-38 for 72 h. Cell viability was determined by detecting the luminescence. E , HCT116-siRNA-CTDSP1 or control siRNA cells were plated in a 6-well plate and treated with various concentrations of SN-38 for 24 h. Then, 50 cells per well were plated in a 6-well plate. After 14 days, when colonies were apparent (right panel), colonies were counted and the relative number of colonies was determined (left panel).

    Article Snippet: ImmunoblottingCells cultured in 6-well plates were scraped into an ice-cold RIPA buffer.

    Techniques: Transfection, CRISPR, Microscopy

    STAT3 nuclear translocation occurs independently of tyrosine phosphorylation. ( A ) HeLa cells (lanes 1 and 2) and HeLa cells transfected with STAT3–GFP (lanes 3 and 4) were untreated (-) or treated (+) with human IL-6 for 30 min. Whole-cell lysates

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

    Article Title: STAT3 nuclear import is independent of tyrosine phosphorylation and mediated by importin-?3

    doi: 10.1073/pnas.0501643102

    Figure Lengend Snippet: STAT3 nuclear translocation occurs independently of tyrosine phosphorylation. ( A ) HeLa cells (lanes 1 and 2) and HeLa cells transfected with STAT3–GFP (lanes 3 and 4) were untreated (-) or treated (+) with human IL-6 for 30 min. Whole-cell lysates

    Article Snippet: Recombinant murine or human IL-6 (BioSource International, Camarillo, CA), IL-6 soluble receptor (R & D Systems) and human EGF (Sigma) were used at 10 ng/ml.

    Techniques: Translocation Assay, Transfection

    Hierarchical clustering of autophagy-related genes in IL-6- and TNF-α-treated placental explants. Human first-trimester placental explants from different patients ( n = 8) were incubated with or without IL-6 (10 ng/ml) and TNF-α (10 ng/ml), respectively, for 48h. RNA from each treatment group was pooled and subjected to autophagy qPCR array. The heat map with dendrograms shows log2 fold-changes in blue for downregulated and in red for upregulated autophagy-related genes in IL-6-treated ( left column ) and TNF-α- treated ( right column ) placental explants relative to untreated controls

    Journal: Histochemistry and Cell Biology

    Article Title: Placental DAPK1 and autophagy marker LC3B-II are dysregulated by TNF-α in a gestational age-dependent manner

    doi: 10.1007/s00418-016-1537-1

    Figure Lengend Snippet: Hierarchical clustering of autophagy-related genes in IL-6- and TNF-α-treated placental explants. Human first-trimester placental explants from different patients ( n = 8) were incubated with or without IL-6 (10 ng/ml) and TNF-α (10 ng/ml), respectively, for 48h. RNA from each treatment group was pooled and subjected to autophagy qPCR array. The heat map with dendrograms shows log2 fold-changes in blue for downregulated and in red for upregulated autophagy-related genes in IL-6-treated ( left column ) and TNF-α- treated ( right column ) placental explants relative to untreated controls

    Article Snippet: For cytokine treatments, complete culture medium was supplemented with 10 ng/ml recombinant human IL-6 (Peprotech) and recombinant human TNF-α (Peprotech), respectively.

    Techniques: Incubation, Real-time Polymerase Chain Reaction

    Expression analysis of CTSD, HDAC6, and DAPK1 in IL-6- and TNF-α-treated placental explants. Human first-trimester placental explants ( n = 8) were incubated with or without IL-6 (10 ng/ml) and TNF-α (10 ng/ml), respectively, for 48h. qPCR analysis was performed for CTSD ( a ), HDAC6 ( b ), and DAPK1 ( c ). Values are given as mean ± SEM. * p

    Journal: Histochemistry and Cell Biology

    Article Title: Placental DAPK1 and autophagy marker LC3B-II are dysregulated by TNF-α in a gestational age-dependent manner

    doi: 10.1007/s00418-016-1537-1

    Figure Lengend Snippet: Expression analysis of CTSD, HDAC6, and DAPK1 in IL-6- and TNF-α-treated placental explants. Human first-trimester placental explants ( n = 8) were incubated with or without IL-6 (10 ng/ml) and TNF-α (10 ng/ml), respectively, for 48h. qPCR analysis was performed for CTSD ( a ), HDAC6 ( b ), and DAPK1 ( c ). Values are given as mean ± SEM. * p

    Article Snippet: For cytokine treatments, complete culture medium was supplemented with 10 ng/ml recombinant human IL-6 (Peprotech) and recombinant human TNF-α (Peprotech), respectively.

    Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction

    PLAG regulated activation of STAT3 A. LPS (100 ng/mL)-treated RAW264.7 cells were treated with Bay 11-7082 (0.1 or 1 μM) or S3I-201 (10 or 100 μM) alone or together with PLAG (1 or 10 μg/mL) for 12 h, and production of IL-6 was analyzed by ELISA of the culture medium. PLAG enhanced the inhibition of IL-6 production by both compounds. * p

    Journal: Oncotarget

    Article Title: 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol ameliorates arthritic joints through reducing neutrophil infiltration mediated by IL-6/STAT3 and MIP-2 activation

    doi: 10.18632/oncotarget.19384

    Figure Lengend Snippet: PLAG regulated activation of STAT3 A. LPS (100 ng/mL)-treated RAW264.7 cells were treated with Bay 11-7082 (0.1 or 1 μM) or S3I-201 (10 or 100 μM) alone or together with PLAG (1 or 10 μg/mL) for 12 h, and production of IL-6 was analyzed by ELISA of the culture medium. PLAG enhanced the inhibition of IL-6 production by both compounds. * p

    Article Snippet: Recombinant mouse or human IL-6 was purchased from PeproTech (Rocky Hill, NJ, USA).

    Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Inhibition

    Pathologic phenotypes of CIA mice were ameliorated by PLAG A. Scheme for generation of CIA mouse model. B. RA was induced by injection of collagen in DBA/1J mice, and mice were examined visually during development of RA and scored based on the severity of their phenotypes. The arthritic phenotypes in CIA mice were ameliorated by PLAG treatment. Ten mice in each group were examined and mean values determined. Values represent mean ± standard error of the mean (SEM). C. Blood was collected from each mouse, and IL-6 levels in plasma were analyzed by ELISA. The increased IL-6 level in the blood was decreased by the administration of PLAG in CIA mice. D. The level of antibodies against type II collagen was not affected by PLAG. Each sample was analyzed in triplicate, and values represent mean ± SEM. # p

    Journal: Oncotarget

    Article Title: 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol ameliorates arthritic joints through reducing neutrophil infiltration mediated by IL-6/STAT3 and MIP-2 activation

    doi: 10.18632/oncotarget.19384

    Figure Lengend Snippet: Pathologic phenotypes of CIA mice were ameliorated by PLAG A. Scheme for generation of CIA mouse model. B. RA was induced by injection of collagen in DBA/1J mice, and mice were examined visually during development of RA and scored based on the severity of their phenotypes. The arthritic phenotypes in CIA mice were ameliorated by PLAG treatment. Ten mice in each group were examined and mean values determined. Values represent mean ± standard error of the mean (SEM). C. Blood was collected from each mouse, and IL-6 levels in plasma were analyzed by ELISA. The increased IL-6 level in the blood was decreased by the administration of PLAG in CIA mice. D. The level of antibodies against type II collagen was not affected by PLAG. Each sample was analyzed in triplicate, and values represent mean ± SEM. # p

    Article Snippet: Recombinant mouse or human IL-6 was purchased from PeproTech (Rocky Hill, NJ, USA).

    Techniques: Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay

    The migration of neutrophils was inhibited by PLAG A. LPS treatment (100 ng/mL) of RA-FLSs increased IL-6 expression, which was inhibited by co-treatment with PLAG in a concentration-dependent manner. B. IL-6 production by LPS-stimulated RA-FLSs was dependent on LPS concentration, and the inhibitory effect of PLAG was significant only after IL-6 production reached a certain level. * p

    Journal: Oncotarget

    Article Title: 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol ameliorates arthritic joints through reducing neutrophil infiltration mediated by IL-6/STAT3 and MIP-2 activation

    doi: 10.18632/oncotarget.19384

    Figure Lengend Snippet: The migration of neutrophils was inhibited by PLAG A. LPS treatment (100 ng/mL) of RA-FLSs increased IL-6 expression, which was inhibited by co-treatment with PLAG in a concentration-dependent manner. B. IL-6 production by LPS-stimulated RA-FLSs was dependent on LPS concentration, and the inhibitory effect of PLAG was significant only after IL-6 production reached a certain level. * p

    Article Snippet: Recombinant mouse or human IL-6 was purchased from PeproTech (Rocky Hill, NJ, USA).

    Techniques: Migration, Expressing, Concentration Assay

    PLAG inhibited IL-6 expression specifically A. RAW264.7 cells were treated with LPS (1 μg/mL) for 12 h, to induce expression of inflammatory cytokines IL-6 and TNF-α and co-treated with the indicated concentration of PLAG. Co-treatment with PLAG inhibited LPS-induced IL-6 expression in a dose-dependent manner. However, PLAG did not affect the expression of TNF-α. B. LPS-treated RAW264.7 cells were analyzed at the indicated times, showing that PLAG inhibited IL-6 transcription in a time-dependent manner. C. Luciferase activity in RAW264.7 cells transfected with pGL4-IL6p was enhanced by treatment with LPS (1 μg/mL). Co-treatment with PLAG inhibited the luciferase activity dose-dependently. * p

    Journal: Oncotarget

    Article Title: 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol ameliorates arthritic joints through reducing neutrophil infiltration mediated by IL-6/STAT3 and MIP-2 activation

    doi: 10.18632/oncotarget.19384

    Figure Lengend Snippet: PLAG inhibited IL-6 expression specifically A. RAW264.7 cells were treated with LPS (1 μg/mL) for 12 h, to induce expression of inflammatory cytokines IL-6 and TNF-α and co-treated with the indicated concentration of PLAG. Co-treatment with PLAG inhibited LPS-induced IL-6 expression in a dose-dependent manner. However, PLAG did not affect the expression of TNF-α. B. LPS-treated RAW264.7 cells were analyzed at the indicated times, showing that PLAG inhibited IL-6 transcription in a time-dependent manner. C. Luciferase activity in RAW264.7 cells transfected with pGL4-IL6p was enhanced by treatment with LPS (1 μg/mL). Co-treatment with PLAG inhibited the luciferase activity dose-dependently. * p

    Article Snippet: Recombinant mouse or human IL-6 was purchased from PeproTech (Rocky Hill, NJ, USA).

    Techniques: Expressing, Concentration Assay, Luciferase, Activity Assay, Transfection

    IL-6 enhances neural stem cell expansion/maintenance, which can be inhibited by COX inhibitors. Primary neurospheres were dissociated and replated into either control media or media supplemented with 5ng/mL of rmIL-6. The number (A) and size (B) of spheres was quantified after 6 days. IL-6 increased the number of secondary neurospheres (*, p

    Journal: Annals of neurology

    Article Title: Opposite Effect Of Inflammation on SVZ vs. Hippocampal Precursors In Brain Injury

    doi: 10.1002/ana.22473

    Figure Lengend Snippet: IL-6 enhances neural stem cell expansion/maintenance, which can be inhibited by COX inhibitors. Primary neurospheres were dissociated and replated into either control media or media supplemented with 5ng/mL of rmIL-6. The number (A) and size (B) of spheres was quantified after 6 days. IL-6 increased the number of secondary neurospheres (*, p

    Article Snippet: Wells were washed, blocked and incubated with either dilutions of rmIL-6 or samples overnight at 4° C. The next day wells were washed and biotinylated anti-IL-6 antibody (R & D Systems) added for 2 hours at 37°C followed by washes and then incubation with AP-conjugated Streptavidin for 1 hour at 37°C.

    Techniques:

    IL-6 induction in mouse macrophage cell line J774A.1 by PagL LPS (A) and LpxL1 LPS (B). Tested were free LPS, liposomes with incorporated LPS, and liposomes with coincorporated LPS and OpaJ at LPS/protein ratios of 1:5 and 1:50 (μmol/μg).

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Coincorporation of LpxL1 and PagL Mutant Lipopolysaccharides into Liposomes with Neisseria meningitidis Opacity Protein: Influence on Endotoxic and Adjuvant Activity ▿

    doi: 10.1128/CVI.00423-09

    Figure Lengend Snippet: IL-6 induction in mouse macrophage cell line J774A.1 by PagL LPS (A) and LpxL1 LPS (B). Tested were free LPS, liposomes with incorporated LPS, and liposomes with coincorporated LPS and OpaJ at LPS/protein ratios of 1:5 and 1:50 (μmol/μg).

    Article Snippet: Cells were incubated for 16 to 18 h at 37°C in a humid atmosphere with 5% CO2 , and the IL-6 concentrations in the supernatants were determined in an enzyme-linked immunosorbent assay (ELISA) against mouse IL-6 according to the manufacturer's instructions (BD Bioscience P, San Diego, CA).

    Techniques:

    MSCs decrease the splenic Th17/Treg ratio and mRNA levels of pro-inflammatory cytokines in the ankle joint. (A, B) CD4, CD25, FoxP3, and RORγt expression in the spleen were determined by flow cytometry. The frequency of Th17 cells was reported as the percentage of CD4+ RORγt+ cells out of the total CD4+ T cell population. The frequency of Treg was reported as the percentage of CD4+CD25+FoxP3+ cells out of the total CD4+ T cell population. (C) Total RNA was isolated from the joints, and the expression of human, mouse IL-1Ra and pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, and IL-17 was investigated. (D-I) The densitometric quantification of the mRNA expression in joints. β-actin expression was used as the endogenous control. Data represent the mean ± SEM. *, P

    Journal: PLoS ONE

    Article Title: Mesenchymal stem cells ameliorate experimental arthritis via expression of interleukin-1 receptor antagonist

    doi: 10.1371/journal.pone.0193086

    Figure Lengend Snippet: MSCs decrease the splenic Th17/Treg ratio and mRNA levels of pro-inflammatory cytokines in the ankle joint. (A, B) CD4, CD25, FoxP3, and RORγt expression in the spleen were determined by flow cytometry. The frequency of Th17 cells was reported as the percentage of CD4+ RORγt+ cells out of the total CD4+ T cell population. The frequency of Treg was reported as the percentage of CD4+CD25+FoxP3+ cells out of the total CD4+ T cell population. (C) Total RNA was isolated from the joints, and the expression of human, mouse IL-1Ra and pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, and IL-17 was investigated. (D-I) The densitometric quantification of the mRNA expression in joints. β-actin expression was used as the endogenous control. Data represent the mean ± SEM. *, P

    Article Snippet: The sorted CD4+ T cells were cultured in RPMI 1640 medium supplemented with 10% FBS (Gibco) and stimulated with 1 μg/ml plate-bound anti-mouse CD3 (BD Biosciences), 2 μg/ml anti-mouse CD28 (BD Biosciences), 2 μg/ml anti-mouse IL-4 (R & D Systems, MN, USA), 2 μg/ml anti-mouse interferon-γ (IFN-γ) (R & D Systems), 20 ng/ml recombinant IL-6 (R & D Systems), and 2 ng/ml recombinant transforming growth factor-β1 (TGF-β1) (R & D Systems) for 3 days.

    Techniques: Expressing, Flow Cytometry, Cytometry, Isolation

    Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic IL-6 levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P

    Journal: Molecular Medicine Reports

    Article Title: Syringaresinol-di-O-β-D-glucoside, a phenolic compound from Polygonatum sibiricum, exhibits an antidiabetic and antioxidative effect on a streptozotocin-induced mouse model of diabetes

    doi: 10.3892/mmr.2018.9580

    Figure Lengend Snippet: Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic IL-6 levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P

    Article Snippet: Insulin (cat. no. ab100578) and interleukin-6 (IL-6; cat. no. ab100712) ELISA kits, and transforming growth factor-β1 (TGF-β1; cat. no. ab9758) and GAPDH (cat. no. ab8245) antibodies, were purchased from Abcam (Cambridge, MA, USA).

    Techniques: Mouse Assay, Injection, Standard Deviation

    Determination of levels of TNF-α and IL-6 in small intestines of AG129 mice after inoculation of ICs. Groups of AG129 mice ( n = 3) were challenged with a sub-lethal dose of DENV-2 S221 (V) or 100% in vitro -neutralized ICs of DENV-2 S221 with mAb 4G2 (V+4G2 ICs), anti-mE1E2 bv VLP antiserum (V+α-mE1E2 bv VLP ICs), anti-pmE1+E2 VLP antiserum (V+ α-pmE1+E2 VLP ICs), anti-mE3E4 bv VLP antiserum (V+α-mE3E4 bv VLP ICs), or anti-DENV-2 S221 antiserum (V+ α-DENV-2 S221 ICs), in parallel with the inoculations described in Figure 6B . One group was included as negative control (NC) which did not receive any treatment. Three days post-inoculation mice were euthanized, small intestines dissected out after perfusion and homogenized. Clarified homogenates were used for the determination of TNF-α (A) and IL-6 (B) using commercial ELISA kits, with purified recombinant murine TNF-α and IL-6 as references. Data shown are the mean values with the bars denoting standard deviation ( n = 3).

    Journal: Frontiers in Microbiology

    Article Title: Pichia pastoris-Expressed Bivalent Virus-Like Particulate Vaccine Induces Domain III-Focused Bivalent Neutralizing Antibodies without Antibody-Dependent Enhancement in Vivo

    doi: 10.3389/fmicb.2017.02644

    Figure Lengend Snippet: Determination of levels of TNF-α and IL-6 in small intestines of AG129 mice after inoculation of ICs. Groups of AG129 mice ( n = 3) were challenged with a sub-lethal dose of DENV-2 S221 (V) or 100% in vitro -neutralized ICs of DENV-2 S221 with mAb 4G2 (V+4G2 ICs), anti-mE1E2 bv VLP antiserum (V+α-mE1E2 bv VLP ICs), anti-pmE1+E2 VLP antiserum (V+ α-pmE1+E2 VLP ICs), anti-mE3E4 bv VLP antiserum (V+α-mE3E4 bv VLP ICs), or anti-DENV-2 S221 antiserum (V+ α-DENV-2 S221 ICs), in parallel with the inoculations described in Figure 6B . One group was included as negative control (NC) which did not receive any treatment. Three days post-inoculation mice were euthanized, small intestines dissected out after perfusion and homogenized. Clarified homogenates were used for the determination of TNF-α (A) and IL-6 (B) using commercial ELISA kits, with purified recombinant murine TNF-α and IL-6 as references. Data shown are the mean values with the bars denoting standard deviation ( n = 3).

    Article Snippet: TNF-α and IL-6 in the clarified small intestinal extracts were determined using commercial cytokine ELISA kits (cat# KMC3011 for TNF-α; cat# KMC0061 for IL-6) procured from Invitrogen as per the manufacturer’s instructions.

    Techniques: Mouse Assay, In Vitro, Negative Control, Enzyme-linked Immunosorbent Assay, Purification, Recombinant, Standard Deviation

    The expression of inflammatory cytokines and fibrotic factors are reduced in STZ-induced diabetic rats. ( A ) The kidney tissues were fixed in formalin and then subjected to immunofluorescence detection of TNF-α (arrow heads pointing to dark-brown dots indicating TNF-α expression). n = 5 per group, ( B ) IL6 protein level ( C ) MCP1 protein level ( D ) TGF-β mRNA level, ( E ) TGF-β protein level with a representative blot, ( F ) Fibronectin (FN) mRNA level in kidney tissues. Data are shown as the means ± SEM *p

    Journal: Aging (Albany NY)

    Article Title: Attenuation of diabetic kidney injury in DPP4-deficient rats; role of GLP-1 on the suppression of AGE formation by inducing glyoxalase 1

    doi: 10.18632/aging.102643

    Figure Lengend Snippet: The expression of inflammatory cytokines and fibrotic factors are reduced in STZ-induced diabetic rats. ( A ) The kidney tissues were fixed in formalin and then subjected to immunofluorescence detection of TNF-α (arrow heads pointing to dark-brown dots indicating TNF-α expression). n = 5 per group, ( B ) IL6 protein level ( C ) MCP1 protein level ( D ) TGF-β mRNA level, ( E ) TGF-β protein level with a representative blot, ( F ) Fibronectin (FN) mRNA level in kidney tissues. Data are shown as the means ± SEM *p

    Article Snippet: The antibodies used were as follows: anti-actin (#8457, Cell Signaling Technology, Danvers, MA, USA), anti-GAPDH (#MAB374, Millipore), anti-RAGE (sc-365154, Santa Cruz Biotechnology, Dallas, TX, USA), anti-DPP4/CD26 (5E8) (sc-8422, Santa Cruz), anti-glyoxalase-1 (sc-101537, Santa Cruz), anti-transforming growth factor (TGF)-β (#3711, Cell Signaling Technology), anti-Lamin A/C (#4777, Cell Signaling Technology, Danvers, MA, USA), anti-nuclear factor-erythroid 2 p45 subunit-related factor-2 (Nrf-2) (ab137550, Abcam), anti-monocyte chemoattractant protein 1 (MCP-1) (ab25124, Abcam), and anti-interleukin 6 (IL-6) (ab9324, Abcam).

    Techniques: Expressing, Immunofluorescence

    Ex-4 treatment reduces MGO-induced inflammatory cytokine expression in rat mesangial cells. Rat mesangial cells were treated either with 1 mM MGO, 10 nM Ex-4, or both for 10 h after synchronization with 1% fetal bovine serum for 13-16 h. ( A ) TNF-α mRNA level, ( B ) MCP-1 mRNA level, ( C ) IL6 mRNA level in rat mesangial cells. Data are shown as the means ± SEM. * p

    Journal: Aging (Albany NY)

    Article Title: Attenuation of diabetic kidney injury in DPP4-deficient rats; role of GLP-1 on the suppression of AGE formation by inducing glyoxalase 1

    doi: 10.18632/aging.102643

    Figure Lengend Snippet: Ex-4 treatment reduces MGO-induced inflammatory cytokine expression in rat mesangial cells. Rat mesangial cells were treated either with 1 mM MGO, 10 nM Ex-4, or both for 10 h after synchronization with 1% fetal bovine serum for 13-16 h. ( A ) TNF-α mRNA level, ( B ) MCP-1 mRNA level, ( C ) IL6 mRNA level in rat mesangial cells. Data are shown as the means ± SEM. * p

    Article Snippet: The antibodies used were as follows: anti-actin (#8457, Cell Signaling Technology, Danvers, MA, USA), anti-GAPDH (#MAB374, Millipore), anti-RAGE (sc-365154, Santa Cruz Biotechnology, Dallas, TX, USA), anti-DPP4/CD26 (5E8) (sc-8422, Santa Cruz), anti-glyoxalase-1 (sc-101537, Santa Cruz), anti-transforming growth factor (TGF)-β (#3711, Cell Signaling Technology), anti-Lamin A/C (#4777, Cell Signaling Technology, Danvers, MA, USA), anti-nuclear factor-erythroid 2 p45 subunit-related factor-2 (Nrf-2) (ab137550, Abcam), anti-monocyte chemoattractant protein 1 (MCP-1) (ab25124, Abcam), and anti-interleukin 6 (IL-6) (ab9324, Abcam).

    Techniques: Expressing

    Immunohistochemistry reactions for analysis of the expression of interleukins (IL-6 and IL-10). (A, B and C), tissues marked with primary antibody anti-interleukin-6 (sc-130326); (D, E and F), tissues marked with primary antibody anti-interleukin-10 (H-160).

    Journal: Acta Ortopedica Brasileira

    Article Title: Extracellular matrix remodeling in experimental intervertebral disc degeneration

    doi: 10.1590/S1413-78522013000300003

    Figure Lengend Snippet: Immunohistochemistry reactions for analysis of the expression of interleukins (IL-6 and IL-10). (A, B and C), tissues marked with primary antibody anti-interleukin-6 (sc-130326); (D, E and F), tissues marked with primary antibody anti-interleukin-10 (H-160).

    Article Snippet: The sections were then incubated overnight at 4°C with the primary antibodies: anti-HPSE1 (HPA1 L-19), anti-HPSE2 (HPA2 C-17), anti-aggrecan (4F4), anti-decorin (N-15), anti-biglycan (L-15), TGFβ1 (sc-146), anti-MMP-9 (H-129), anti-interleukin-6 (SC-130326) and anti-interleukin-10 (H-160) (Santa Cruz Biotechnology, CA, USA).

    Techniques: Immunohistochemistry, Expressing

    Supplementation with  Lactobacillus reuteri  100-23 and  Lactobacillus gasseri  311476 mixture reduces inflammatory cytokines. A–F . Plasma levels of interleukin 4 (IL-4), interleukin 10 (IL-10), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8), and granulocyte colony-stimulating factor (G-CSF) in control mice (CT), in mice receiving lactobacilli (Lrg), in mice transplanted with BaF3 cells (BaF3) and in mice transplanted with BaF3 cells and receiving lactobacilli (BaF3-Lrg). Data with different superscript letters are significantly different (p

    Journal: PLoS ONE

    Article Title: Restoring Specific Lactobacilli Levels Decreases Inflammation and Muscle Atrophy Markers in an Acute Leukemia Mouse Model

    doi: 10.1371/journal.pone.0037971

    Figure Lengend Snippet: Supplementation with Lactobacillus reuteri 100-23 and Lactobacillus gasseri 311476 mixture reduces inflammatory cytokines. A–F . Plasma levels of interleukin 4 (IL-4), interleukin 10 (IL-10), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8), and granulocyte colony-stimulating factor (G-CSF) in control mice (CT), in mice receiving lactobacilli (Lrg), in mice transplanted with BaF3 cells (BaF3) and in mice transplanted with BaF3 cells and receiving lactobacilli (BaF3-Lrg). Data with different superscript letters are significantly different (p

    Article Snippet: Plasma interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8) and granulocyte colony-stimulating factor (G-CSF) were measured using a customized multiplex kit (Bio-Rad, Nazareth, Belgium) with the Luminex technology (Bio-Plex, Bio-Rad).

    Techniques: Mouse Assay

    Arsenic induced miR-301a expression dependent on IL-6/STAT3 signaling. (A) BEAS-2B cells were treated with arsenic (5 and 10 μM) for 24 h and with H2O2 as positive control. The levels of IL-6 in the culture medium was determined by using ELISA Kit. (B) BEAS-2B cells were treated with different concentrations of IL-6 (0, 10, 50, and 100 ng/mL) and IL-6 plus Stat3 inhibitor (S31-201, 30 μM), at 6 h and the expression of miR-301a was determined by qPCR. (C) BEAS-2B cells were treated with arsenic (As, 10 μM), arsenic (As) and S31-201, arsenic (As) and IL-6 neutralizing antibody (anti-IL-6) at 6 h and the expression of miR-301a was determined by qPCR. (D) BEAS-2B cells were transfected with Anti-control (Anti-control) and LNA-Anti-miR-301a(Anti-miR-301a). After 48 h post transfection, cells were treated with arsenic (5 μM and 10 μM) for 24 h and western blot analyses of phospho-Stat3 (pStat3), Stat3, IκBα expression. Gel pictures were acquired by using Tanon5200 and the exposure time was 60 sec for pStat3, 20 sec for Stat3, 20 sec for IκBα, and 5 sec for β-actin. (E) Protein expression from (D) were quantified by band intensity and normalized to β-actin. ( F ) pStat3 expression was quantified by band intensity and normalized to total Stat3. Values represented the mean ± SD of three independent experiments. *P

    Journal: Scientific Reports

    Article Title: Malignant Transformation of Human Bronchial Epithelial Cells Induced by Arsenic through STAT3/miR-301a/SMAD4 Loop

    doi: 10.1038/s41598-018-31516-0

    Figure Lengend Snippet: Arsenic induced miR-301a expression dependent on IL-6/STAT3 signaling. (A) BEAS-2B cells were treated with arsenic (5 and 10 μM) for 24 h and with H2O2 as positive control. The levels of IL-6 in the culture medium was determined by using ELISA Kit. (B) BEAS-2B cells were treated with different concentrations of IL-6 (0, 10, 50, and 100 ng/mL) and IL-6 plus Stat3 inhibitor (S31-201, 30 μM), at 6 h and the expression of miR-301a was determined by qPCR. (C) BEAS-2B cells were treated with arsenic (As, 10 μM), arsenic (As) and S31-201, arsenic (As) and IL-6 neutralizing antibody (anti-IL-6) at 6 h and the expression of miR-301a was determined by qPCR. (D) BEAS-2B cells were transfected with Anti-control (Anti-control) and LNA-Anti-miR-301a(Anti-miR-301a). After 48 h post transfection, cells were treated with arsenic (5 μM and 10 μM) for 24 h and western blot analyses of phospho-Stat3 (pStat3), Stat3, IκBα expression. Gel pictures were acquired by using Tanon5200 and the exposure time was 60 sec for pStat3, 20 sec for Stat3, 20 sec for IκBα, and 5 sec for β-actin. (E) Protein expression from (D) were quantified by band intensity and normalized to β-actin. ( F ) pStat3 expression was quantified by band intensity and normalized to total Stat3. Values represented the mean ± SD of three independent experiments. *P

    Article Snippet: Human recombinant IL-6 was purchased from R & D Systems (R & D, Minneapolis, MN).

    Techniques: Expressing, Positive Control, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Size-exclusion Chromatography

    a - f Mitochondrial proteins and inflammatory cytokines levels in explant cultured media in SIM tissue and surrounding matched-normal tissue. Wilcoxon matched-pairs signed rank tests demonstrated significantly increased levels of a cytochrome c (n=12), b SMAC/Diablo (n=8), c IL-1beta (n=12), d IL-6 (n=12), e IL-8 (n=12) and f TNF-alpha (n=12) in SIM tissue compared to surrounding normal epithelium. * p ≤ 0.05, ** p ≤ 0.005 and *** p ≤ 0.0005

    Journal: BMC Cancer

    Article Title: Changes in mitochondrial stability during the progression of the Barrett’s esophagus disease sequence

    doi: 10.1186/s12885-016-2544-2

    Figure Lengend Snippet: a - f Mitochondrial proteins and inflammatory cytokines levels in explant cultured media in SIM tissue and surrounding matched-normal tissue. Wilcoxon matched-pairs signed rank tests demonstrated significantly increased levels of a cytochrome c (n=12), b SMAC/Diablo (n=8), c IL-1beta (n=12), d IL-6 (n=12), e IL-8 (n=12) and f TNF-alpha (n=12) in SIM tissue compared to surrounding normal epithelium. * p ≤ 0.05, ** p ≤ 0.005 and *** p ≤ 0.0005

    Article Snippet: The levels of cytokines interleukin-8 (IL-8), interleukin-6 (IL-6), interleukin-1β (IL-1β) and tumour necrosis factor-α (TNF-α) were measured using a multiplex assay from Mesoscale Discovery® (Gaithersburg, MD, USA) using protocols as per manufactures’ instructions.

    Techniques: Cell Culture