mcp-1 Search Results


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  • 95
    Millipore mcp 1
    Human mesenchymal stem cells express lower levels of proinflammatory cytokines when cultured with the novel primary wound dressings. Human mesenchymal stem cells were cultured in the presence or absence of dressing C or A for 4 days in the presence or absence of TGFβ before supernatants were collected and analyzed by Luminex for production of the inflammatory cytokines, IL-6 (a), <t>MCP-1</t> (b), IL-8 (c), and GRO, also called CXCL1 (d). The assay was performed in triplicate, and data are presented as mean ± SEM ( * P
    Mcp 1, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1274 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mcp 1  (Abcam)
    92
    Abcam mcp 1
    Histological and immunohistochemical characteristics of the IEA. The intima thickening is composed of SMA-positive cells (magnification ×200) ( A ) and collagen fibers positive for Sirius Red (magnification ×100) ( B ). Macrophages only are detectable as isolated cells in the intima (magnification ×400) ( C ) or forming aggregates around medial calcifications (magnification ×100) ( D ). Abundant localization of <t>MCP-1–positive</t> cells ( E ), as well as IL-6–positive cells ( F ), is present in both the intima and muscular layers. Strong VCAM-1 expression is observed in the endothelial layer ( G ) of the artery wall (magnification ×400). Note that endothelial and SMCs, as well as SMA-positive cells in the neointima, are positive for the three molecules. A and B represent contiguous sections of the same artery; note the fibrous cap formed by SMA-positive cells and collagen. Arrow, internal elastic lamina; L, lumen. (A high-quality digital representation of this figure is available in the online issue.)
    Mcp 1, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 1195 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology mcp 1
    Effects of STZ and GH treatment on renal MCP immunoreactivity. ( A ) STZ increases the <t>MCP-1</t> staining in kidneys from diabetic male and female rats; however, GH treatment increased MCP-1 only in kidneys from diabetic male rats. * P
    Mcp 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1001 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher mcp 1
    Enhanced chemoattractant activity in CAV-1-silenced monocytes. The chemoattractant activity of monocytes toward <t>MCP-1</t> was assessed by chemotaxis assay. Cav-1-silenced (siCAV-1) or control monocytes (siNeg) were seeded in the upper compartment, and the medium containing 100 ng/ml MCP-1 was then added to the lower compartment. After a 90-minute incubation, cells were collected from both compartments and manually counted. Graph displays the ratio of migrating monocytes. Monocytes were obtained from three donors, and results represented three independent experiments. Data are shown as a mean ± SD. One-way analysis of variance (post hoc Tukey), ***P
    Mcp 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 2982 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Pharmingen mcp 1
    a, Numerous <t>MCP-1+</t> capillaries in a group 1 tissue section from a periodontitis patient demonstrating little inflammatory activity (original magnification ×200). b, Same area of section showing no MIP-1alpha+ endothelial cells.
    Mcp 1, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 295 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    RayBiotech mcp 1
    Summary diagram . TLR4 or TNFα receptor stimulation increases the activity of NF-κB and the expression of its target genes NR4A1–3 (NR4A), <t>MCP-1,</t> MIP-3α, and Relb. Loss of NR4A potentiates this induction of MCP-1, while reducing the induction of Relb and MIP-3α. Furthermore using mouse embryonic cells, we identify MIP-3α is a novel Relb target gene. Therefore, NR4A receptors act as repressors of inflammatory-NF-κB-driven MCP-1 (indicated by a red T bar) and enhancers of NF-κB-driven Relb and subsequently MIP-3α. Using a NR4A2 DNA-binding mutant reveals that NR4A2 does not require the ability to bind DNA in order to enhance/repress target genes simultaneously.
    Mcp 1, supplied by RayBiotech, used in various techniques. Bioz Stars score: 93/100, based on 291 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Becton Dickinson mcp 1
    Quantitative chemokine mRNA expression in the CNS. The relative amount of mRNA transcripts for (A) <t>MCP-1</t> and (B) MIP-1α was analyzed from spinal cord tissue of MBP-EAE (MBP), S100β-EAE (S100), and control animals (control) by dot blot hybridization at days 2 and 4 after transfer. Note the massive up-regulation of MCP-1 and MIP-1α mRNA in the CNS of T MBP cell–treated, but not in the T S100β cell–treated, animals. Amplification of β-tubulin and analysis on ethidium bromide–stained agarose gels demonstrated intact RNA in all samples (not depicted). The data were confirmed in a second set of independently prepared samples.
    Mcp 1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 2425 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cell Signaling Technology Inc mcp 1
    MPs are released by stressed adipocytes in a caspase 3 and Rho-associated kinase dependent manner. (A-D) Characterization of adipocyte-derived MPs. (A) Morphology of the differentiated 3T3-L1 adipocytes treated with control, 0.5 mM palmitic acid. (B) The number of Annexin V positive MPs was quantitated by flow cytometry. (C) Dynamic light scattering analysis and Transmission Electron Microscopy of isolated MPs. Isolated MPs were measured by Zetasizer and analyzed using intensity. (D) Western Blot analysis of MPs released by adipocytes. Isolated MPs were fractionated by SDS-PAGE and probed with FABP4, <t>MCP-1,</t> Chemerin, Adiponectin and Perilipin antibody. (E-F) Differentiated adipocytes were incubated with or without palmitic acid in the absence or presence of a selective caspase-3 inhibitor (E), or a range of doses of two different Rho associated kinase inhibitors (Y27632 and fasudil) (F) for up to 12 hrs. Supernatants were then collected and MPs isolated by ultracentrifugation as detailed in methods section. (G) Morphology of isolated mouse primary adipocytes. (H) Number of annexin V positive mouse primary adipocyte-derived MPs assessed by flow cytometry. Values represent mean ± S.D. * P
    Mcp 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 310 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Promega mcp 1
    Effect of <t>MCP-1</t> on chemotaxis of CCR2-positive mononuclear cells in viral myocarditis in vitro. Peripheral blood mononuclear cells were separated from mice 7 days post-CVB3 infection. The purified peripheral blood mononuclear cells were analyzed for the
    Mcp 1, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    The Jackson Laboratory mcp 1
    <t>MCP-1</t> and survival
    Mcp 1, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 93/100, based on 112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam anti mcp 1
    The effect of STAMP2 gene overexpression on inflammatory factors expression in WAT and BAT of ApoE −/− /LDLR −/− mice. A: Immunohistochemical staining for adipose tissue IL-6, TNF-α, <t>MCP-1</t> and IL-10 (brown staining considered positive staining; scale bar: 50 µm). B: Semiquantification of IL-6, TNF-α, MCP-1 and IL-10 immunohistochemical staining. Data are mean ± SEM (three sections per mice; n = 4 per group). * P
    Anti Mcp 1, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 227 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    PeproTech mcp 1
    A physiologically relevant dose of the formulation can inhibit <t>MCP-1</t> induced migration of human monocytes. Cellular migration was assessed using THP-1 monocytes that were treated with vehicle (vehicle control) or treated with MCP-1 (20 ng/ml) alone or with MCP-1 (20 ng/ml) in the presence the complete formulation (+PS) or with MCP-1 (20ng/ml) in the presence the formulation lacking PS (-PS) for 3 hours. Monocyte migration is expressed as a fold-change compared to the proportion of cells that moved from the apical compartment into the basolateral compartment in response to MCP-1 alone that has been arbitrarily set as 1. The data are presented as the mean ± SEM from four independent experiments. Statistical analysis was performed using a one-way ANOVA with Dunnett T3 post-hoc analysis where ** p
    Mcp 1, supplied by PeproTech, used in various techniques. Bioz Stars score: 97/100, based on 847 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti mcp1 antibody
    Transplantation of rat NPMSCs overexpressing SIRT1 relieves IDD in rats by downregulating the <t>MCP1/CCR2</t> axis. (A) RT-qPCR and (B) western blot analyses showed increased mRNA and protein expression of MCP1 and CCR2, and reduced mRNA and protein expression of SIRT1 in the IDD group compared with the control group. In contrast, the mRNA and protein expression of MCP1 and CCR2 decreased and mRNA and protein expression of SIRT1 increased in the IDD rats after transplantation of the NPMSCs or SIRT1-overexpressing NPMSCs, n=4. (C) H E staining revealed an amelioration of the disordered intervertebral structure after the transplantation of NPMSCs or SIRT1-overexpressing NPMSCs, n=6. Based on the (D) RT-qPCR and (E) western blot analyses, lower mRNA and protein expression levels of aggrecan, collagen II and Sox-9 were observed in IDD rats compared with in rats of the control group, n=4. (F) TUNEL staining revealed increased cell apoptosis in IDD rats compared with rats in the control group, which was reduced by NPMSC transplantation and even further decreased after the transplantation of SIRT1-overexpressing NPMSCs, n=6. (G) Western blotting showed higher protein expression of Bax and cleaved caspase-3 and lower expression of Bcl-2 in the IDD group compared with in the control group, and these changes were reversed by the transplantation of NPMSCs or SIRT1-overexpressing NPMSCs, n=4. Data in panel F were analyzed using one-way ANOVA; data in panels A, B, C, D, E and G were analyzed using two-way ANOVA. Tukey's multiple comparisons test was applied as the post hoc test. ** P
    Anti Mcp1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 116 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    USCN Life mcp 1
    AS-IV decreased the serum levels of TNF- α , <t>MCP-1,</t> and ICAM-1 in rats with ischemic AKI. Serum levels of TNF- α (a), MCP-1 (b), and ICAM-1 (c) in sham, vehicle-, or AS-IV-pretreated rats at 12 h of reperfusion. Serum levels of TNF- α (d), MCP-1 (e), and ICAM-1 (f) in sham, vehicle-, or AS-IV-pretreated rats at 24 h of reperfusion. Results are expressed as mean ± SD ( n = 8). * P
    Mcp 1, supplied by USCN Life, used in various techniques. Bioz Stars score: 92/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    R&D Systems recombinant human mcp 1
    Monocyte chemotactic activity in supernatants of endothelial cells. Monolayers of endothelial cells were stimulated with 100 U/ml recombinant human IL-β, or incubated with 5 × 10 7 strain 42D or with 1 × 10 7 strain CAPD for 60 min at 37°C. After washing and lysostaphin treatment, the endothelial cells were subsequently cultured. At different time points, culture supernatants were harvested and tested for chemotactic activity towards human monocytes. A representative experiment is shown. Data are expressed as the ratio between the number of migrated monocytes towards culture supernatant of infected or IL-1-β-stimulated endothelial cells and the number of migrated monocytes towards culture supernatant of untreated endothelial cells. Chemotaxis was tested in the presence (□) or absence (▪) of neutralizing <t>anti-MCP-1</t> antibody.
    Recombinant Human Mcp 1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    MyBiosource mcp 1
    IVC levels of TNF-α, IF-γ, <t>MCP-1</t> and IL-10 in normal, RAS and RAS + MSC pigs ( A – D ). Urine levels of the same cytokines did not differ among the groups ( E – H ). Net release of TNF-α, IF-γ, MCP-1 and IL-10 in
    Mcp 1, supplied by MyBiosource, used in various techniques. Bioz Stars score: 92/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology anti mcp 1
    Emodin reduces E-selectin and <t>MCP-1</t> expression in the lung tissues in LPS-induced ALI. After 72 h interventions, mice were exsanguinated and their lungs were removed. ( A , C ) qPCR was performed to analyze E-selectin and MCP-1 mRNA expression in the lung tissues; ( B , D ) Western blotting was performed to measure E-selectin and MCP-1 protein expression in the lung tissues. Each bar represents the mean ± SD of 10 mice. * p
    Anti Mcp 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 178 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Randox mcp 1
    Emodin reduces E-selectin and <t>MCP-1</t> expression in the lung tissues in LPS-induced ALI. After 72 h interventions, mice were exsanguinated and their lungs were removed. ( A , C ) qPCR was performed to analyze E-selectin and MCP-1 mRNA expression in the lung tissues; ( B , D ) Western blotting was performed to measure E-selectin and MCP-1 protein expression in the lung tissues. Each bar represents the mean ± SD of 10 mice. * p
    Mcp 1, supplied by Randox, used in various techniques. Bioz Stars score: 92/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson monocyte chemoattractant protein mcp 1
    Release of cytokines by purified natural killer (NK) cells upon stimulation with selected expansion protocols. Purified NK cells were cultured using different stimulation protocols. On day 6, release of tumor necrosis factor (TNF)-α, IFN-γ, macrophage inflammatory protein (MIP)-1α, monocyte <t>chemoattractant</t> protein <t>(MCP)-1,</t> granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-8, and IL-10 were assayed by cytometric bead array analyses. Graphs show mean values and SDs obtained from four independent donors, except GM-CSF, which was measured for three independent donors. Asterisks indicate significant differences (* p
    Monocyte Chemoattractant Protein Mcp 1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 160 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems recombinant mouse ccl2
    Activation of the FoxO1–KLF2–S1P1 axis by CCR2 signaling through Stat3. (A) Surface expression of S1P1 in the CD69 + Qa-2 − (SP1 + SP2), CD69 − Qa-2 − (SP3), and CD69 − Qa-2 + (SP4) subsets of CD4 SP thymocytes from WT and KO mice. The experiments were repeated three times with six mice for each group. Representative dot plots are shown. (B) Percentage of S1P1 + cells are presented as mean ± SD. (C) Foxo1, Klf2, and S1pr1 mRNA expression in WT and KO CD4 SP thymocytes as determined by qPCR. The experiments were repeated three times with triplicates. Data are present as mean ± SEM. (D) CD69 − Qa-2 + SP4 thymocytes from WT and KO mice were stimulated with <t>CCL2</t> in the presence or absence of STAT3 inhibitor Stattic. Foxo1, Klf2, and S1pr1 mRNA expression was detected by qPCR. The experiments were repeated three times with triplicates. Data are present as mean ± SEM. (E) Following stimulation with CCL2, WT SP4 thymocytes were examined for surface expression of S1P1. Representative histograms are shown out of three independent experiments. (F,G) Levels of phosphorylated Stat3 were examined by Western blotting (F) or intracellular staining (G) in SP4 thymocytes after stimulation with CCL2. Western blotting was performed at 30 min after stimulation. The experiments were repeated three times with similar results. (H–J) SP4 thymocytes were stimulated with CCL2 in the presence or absence of STAT3 inhibitor Stattic. Surface expression of S1P1 was assayed by flow cytometry (H) . Subcellular localization of FoxO1 was revealed by immunofluorescent staining (I) and nucleus Foxo1 was quantified by mean fluorescent intensity (MFI) (J) . Data are presented as mean ± SD. Two independent experiments were performed with similar results. * p
    Recombinant Mouse Ccl2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad mcp 1
    miR-K10a-mediated downregulation of TWEAKR inhibits production of IL-8 and <t>MCP-1.</t> (A) HUVECs were transfected with miR-K10a and treated with TWEAK at 24 hpt. Culture supernatants were harvested at 24 h posttreatment and analyzed for IL-8 (top) and MCP-1
    Mcp 1, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1058 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Human mesenchymal stem cells express lower levels of proinflammatory cytokines when cultured with the novel primary wound dressings. Human mesenchymal stem cells were cultured in the presence or absence of dressing C or A for 4 days in the presence or absence of TGFβ before supernatants were collected and analyzed by Luminex for production of the inflammatory cytokines, IL-6 (a), MCP-1 (b), IL-8 (c), and GRO, also called CXCL1 (d). The assay was performed in triplicate, and data are presented as mean ± SEM ( * P

    Journal: Eplasty

    Article Title: In Vitro Characterization of Variable Porosity Wound Dressing With Anti-Scar Properties

    doi:

    Figure Lengend Snippet: Human mesenchymal stem cells express lower levels of proinflammatory cytokines when cultured with the novel primary wound dressings. Human mesenchymal stem cells were cultured in the presence or absence of dressing C or A for 4 days in the presence or absence of TGFβ before supernatants were collected and analyzed by Luminex for production of the inflammatory cytokines, IL-6 (a), MCP-1 (b), IL-8 (c), and GRO, also called CXCL1 (d). The assay was performed in triplicate, and data are presented as mean ± SEM ( * P

    Article Snippet: Conditioned culture medium was collected and analyzed for IL-6, MCP-1, IL-8, and GRO.

    Techniques: Cell Culture, Luminex

    Histological and immunohistochemical characteristics of the IEA. The intima thickening is composed of SMA-positive cells (magnification ×200) ( A ) and collagen fibers positive for Sirius Red (magnification ×100) ( B ). Macrophages only are detectable as isolated cells in the intima (magnification ×400) ( C ) or forming aggregates around medial calcifications (magnification ×100) ( D ). Abundant localization of MCP-1–positive cells ( E ), as well as IL-6–positive cells ( F ), is present in both the intima and muscular layers. Strong VCAM-1 expression is observed in the endothelial layer ( G ) of the artery wall (magnification ×400). Note that endothelial and SMCs, as well as SMA-positive cells in the neointima, are positive for the three molecules. A and B represent contiguous sections of the same artery; note the fibrous cap formed by SMA-positive cells and collagen. Arrow, internal elastic lamina; L, lumen. (A high-quality digital representation of this figure is available in the online issue.)

    Journal: Diabetes Care

    Article Title: Type 1 Diabetes Increases the Expression of Proinflammatory Cytokines and Adhesion Molecules in the Artery Wall of Candidate Patients for Kidney Transplantation

    doi: 10.2337/dc11-1665

    Figure Lengend Snippet: Histological and immunohistochemical characteristics of the IEA. The intima thickening is composed of SMA-positive cells (magnification ×200) ( A ) and collagen fibers positive for Sirius Red (magnification ×100) ( B ). Macrophages only are detectable as isolated cells in the intima (magnification ×400) ( C ) or forming aggregates around medial calcifications (magnification ×100) ( D ). Abundant localization of MCP-1–positive cells ( E ), as well as IL-6–positive cells ( F ), is present in both the intima and muscular layers. Strong VCAM-1 expression is observed in the endothelial layer ( G ) of the artery wall (magnification ×400). Note that endothelial and SMCs, as well as SMA-positive cells in the neointima, are positive for the three molecules. A and B represent contiguous sections of the same artery; note the fibrous cap formed by SMA-positive cells and collagen. Arrow, internal elastic lamina; L, lumen. (A high-quality digital representation of this figure is available in the online issue.)

    Article Snippet: Indeed, a higher expression of IL-6, MCP-1, and adhesion molecules was observed in the artery wall of type 1 diabetic patients compared with nondiabetic subjects.

    Techniques: Immunohistochemistry, Isolation, Expressing

    ANE under lower serum condition induces inflammatory cytokines more efficiently. (A) OC2 cells cultured with indicated concentrations of FBS were treated with 0.5 mg/ml ANE. The cells were harvested for measurement of IL6, IL8, RANTES or VEGF transcripts 24 h later. The value of the untreated, 10% FBS-supplemented cells was defined as 1. (B) The cells transfected with IL8 or COX2 promoter reporter were cultured in medium containing 1% FBS. The luciferase activity was determined 24 h after ANE treatment. (C) Similarly treated cells were harvested for Western blot. The upper signal marked with asterisk might be a glycosylated form of MCP1.

    Journal: Toxicology Reports

    Article Title: Areca nut extracts exert different effects in oral cancer cells depending on serum concentration: A clue to the various oral alterations in betel quid chewers

    doi: 10.1016/j.toxrep.2014.10.018

    Figure Lengend Snippet: ANE under lower serum condition induces inflammatory cytokines more efficiently. (A) OC2 cells cultured with indicated concentrations of FBS were treated with 0.5 mg/ml ANE. The cells were harvested for measurement of IL6, IL8, RANTES or VEGF transcripts 24 h later. The value of the untreated, 10% FBS-supplemented cells was defined as 1. (B) The cells transfected with IL8 or COX2 promoter reporter were cultured in medium containing 1% FBS. The luciferase activity was determined 24 h after ANE treatment. (C) Similarly treated cells were harvested for Western blot. The upper signal marked with asterisk might be a glycosylated form of MCP1.

    Article Snippet: However, it is possible that ANE enhanced deglycosylation rather than expression of MCP1 since Western blotting showed that the increase of MCP1 after ANE treatment was correlated with significant reduction of a high-molecular-weight form of MCP1.

    Techniques: Cell Culture, Transfection, Luciferase, Activity Assay, Western Blot

    Effects of STZ and GH treatment on renal MCP immunoreactivity. ( A ) STZ increases the MCP-1 staining in kidneys from diabetic male and female rats; however, GH treatment increased MCP-1 only in kidneys from diabetic male rats. * P

    Journal: Biology of Sex Differences

    Article Title: Growth hormone exacerbates diabetic renal damage in male but not female rats

    doi: 10.1186/2042-6410-4-12

    Figure Lengend Snippet: Effects of STZ and GH treatment on renal MCP immunoreactivity. ( A ) STZ increases the MCP-1 staining in kidneys from diabetic male and female rats; however, GH treatment increased MCP-1 only in kidneys from diabetic male rats. * P

    Article Snippet: While GH had no additional effect in female rats, there were 3.5 times more MCP-1+ cells in kidneys from STZ + GH male rats when compared to STZ alone (Figure A).

    Techniques: Staining

    Monocyte chemotactic protein 1 ( MCP1 ) is increased in abdominal aortic aneurysm ( AAA ) and required for disease progression. A, Immunohistochemistry performed 14 days after surgery indicates MCP1 is upregulated in elastase-perfused AAAs compared with saline-perfused

    Journal: The Journal of thoracic and cardiovascular surgery

    Article Title: Bone marrow\u2013derived MCP1 required for experimental aortic aneurysm formation and smooth muscle phenotypic modulation

    doi: 10.1016/j.jtcvs.2011.07.053

    Figure Lengend Snippet: Monocyte chemotactic protein 1 ( MCP1 ) is increased in abdominal aortic aneurysm ( AAA ) and required for disease progression. A, Immunohistochemistry performed 14 days after surgery indicates MCP1 is upregulated in elastase-perfused AAAs compared with saline-perfused

    Article Snippet: It is therefore uncertain whether MCP1 is predominantly produced by bone marrow–derived cells or vessel wall resident cells.

    Techniques: Immunohistochemistry

    Mice lacking monocyte chemotactic protein 1 ( MCP1 ) in bone marrow do not form aneurysms after elastase perfusion. Wild type ( WT ) mice with bone marrow cells ( BMCs ) donated from MCP1KO mice (MCP1KO→WT) showed significant reduction in aortic dilation

    Journal: The Journal of thoracic and cardiovascular surgery

    Article Title: Bone marrow\u2013derived MCP1 required for experimental aortic aneurysm formation and smooth muscle phenotypic modulation

    doi: 10.1016/j.jtcvs.2011.07.053

    Figure Lengend Snippet: Mice lacking monocyte chemotactic protein 1 ( MCP1 ) in bone marrow do not form aneurysms after elastase perfusion. Wild type ( WT ) mice with bone marrow cells ( BMCs ) donated from MCP1KO mice (MCP1KO→WT) showed significant reduction in aortic dilation

    Article Snippet: It is therefore uncertain whether MCP1 is predominantly produced by bone marrow–derived cells or vessel wall resident cells.

    Techniques: Mouse Assay

    Knockout of monocyte chemotactic protein 1 ( MCP1 ) in bone marrow cells ( BMCs ) decreases inflammatory markers and increases intact elastic fibers and SMαA expression. A, Immunohistochemistry performed 14 days after elastase perfusion indicates

    Journal: The Journal of thoracic and cardiovascular surgery

    Article Title: Bone marrow\u2013derived MCP1 required for experimental aortic aneurysm formation and smooth muscle phenotypic modulation

    doi: 10.1016/j.jtcvs.2011.07.053

    Figure Lengend Snippet: Knockout of monocyte chemotactic protein 1 ( MCP1 ) in bone marrow cells ( BMCs ) decreases inflammatory markers and increases intact elastic fibers and SMαA expression. A, Immunohistochemistry performed 14 days after elastase perfusion indicates

    Article Snippet: It is therefore uncertain whether MCP1 is predominantly produced by bone marrow–derived cells or vessel wall resident cells.

    Techniques: Knock-Out, Expressing, Immunohistochemistry

    Monocyte chemotactic protein 1 ( MCP1 ) acts on cultured aortic smooth muscle cells ( SMCs ) to repress SMC marker genes and induce matrix metalloproteinases ( MMPs ). Cells treated with 0 to 50 ng/mL MCP1 showed a dose-dependent increase in (A) MMP2 and (B)

    Journal: The Journal of thoracic and cardiovascular surgery

    Article Title: Bone marrow\u2013derived MCP1 required for experimental aortic aneurysm formation and smooth muscle phenotypic modulation

    doi: 10.1016/j.jtcvs.2011.07.053

    Figure Lengend Snippet: Monocyte chemotactic protein 1 ( MCP1 ) acts on cultured aortic smooth muscle cells ( SMCs ) to repress SMC marker genes and induce matrix metalloproteinases ( MMPs ). Cells treated with 0 to 50 ng/mL MCP1 showed a dose-dependent increase in (A) MMP2 and (B)

    Article Snippet: It is therefore uncertain whether MCP1 is predominantly produced by bone marrow–derived cells or vessel wall resident cells.

    Techniques: Cell Culture, Marker

    Enhanced chemoattractant activity in CAV-1-silenced monocytes. The chemoattractant activity of monocytes toward MCP-1 was assessed by chemotaxis assay. Cav-1-silenced (siCAV-1) or control monocytes (siNeg) were seeded in the upper compartment, and the medium containing 100 ng/ml MCP-1 was then added to the lower compartment. After a 90-minute incubation, cells were collected from both compartments and manually counted. Graph displays the ratio of migrating monocytes. Monocytes were obtained from three donors, and results represented three independent experiments. Data are shown as a mean ± SD. One-way analysis of variance (post hoc Tukey), ***P

    Journal: Scientific Reports

    Article Title: Downregulated Caveolin-1 expression in circulating monocytes may contribute to the pathogenesis of psoriasis

    doi: 10.1038/s41598-018-36767-5

    Figure Lengend Snippet: Enhanced chemoattractant activity in CAV-1-silenced monocytes. The chemoattractant activity of monocytes toward MCP-1 was assessed by chemotaxis assay. Cav-1-silenced (siCAV-1) or control monocytes (siNeg) were seeded in the upper compartment, and the medium containing 100 ng/ml MCP-1 was then added to the lower compartment. After a 90-minute incubation, cells were collected from both compartments and manually counted. Graph displays the ratio of migrating monocytes. Monocytes were obtained from three donors, and results represented three independent experiments. Data are shown as a mean ± SD. One-way analysis of variance (post hoc Tukey), ***P

    Article Snippet: RPMI-1640 supplemented with 1% FBS in either the presence or absence of 100 ng/ml MCP-1 (Thermo Fisher Scientific) was added into the lower compartment.

    Techniques: Activity Assay, Chemotaxis Assay, Incubation

    a, Numerous MCP-1+ capillaries in a group 1 tissue section from a periodontitis patient demonstrating little inflammatory activity (original magnification ×200). b, Same area of section showing no MIP-1alpha+ endothelial cells.

    Journal: Clinical and Experimental Immunology

    Article Title: Chemokines in human periodontal disease tissues

    doi: 10.1046/j.1365-2249.2001.01511.x

    Figure Lengend Snippet: a, Numerous MCP-1+ capillaries in a group 1 tissue section from a periodontitis patient demonstrating little inflammatory activity (original magnification ×200). b, Same area of section showing no MIP-1alpha+ endothelial cells.

    Article Snippet: Although the percent MCP-1+ and RANTES+ cells appeared to decrease with increasing inflammation in periodontitis and healthy/gingivitis tissues, respectively, these differences were not significant.

    Techniques: Activity Assay

    The mean (± standard error of the mean) percentage IP-10+ (▪), RANTES+ ( ), MCP-1+ ( ) and MIP-1alpha+ (□) cells in the inflammatory infiltrates of gingival lesions (graded on the size of infiltrate as small in group 1, intermediate in group 2 and extensive in group 3) from healthy/gingivitis (H/G) subjects.

    Journal: Clinical and Experimental Immunology

    Article Title: Chemokines in human periodontal disease tissues

    doi: 10.1046/j.1365-2249.2001.01511.x

    Figure Lengend Snippet: The mean (± standard error of the mean) percentage IP-10+ (▪), RANTES+ ( ), MCP-1+ ( ) and MIP-1alpha+ (□) cells in the inflammatory infiltrates of gingival lesions (graded on the size of infiltrate as small in group 1, intermediate in group 2 and extensive in group 3) from healthy/gingivitis (H/G) subjects.

    Article Snippet: Although the percent MCP-1+ and RANTES+ cells appeared to decrease with increasing inflammation in periodontitis and healthy/gingivitis tissues, respectively, these differences were not significant.

    Techniques:

    The mean (± standard error of the mean) percentage IP-10+ (▪), RANTES+ ( ), MCP-1+ ( ) and MIP-1alpha+ (□) cells in the inflammatory infiltrates of gingival lesions (graded on the size of infiltrate as small in group 1, intermediate in group 2 and extensive in group 3) from periodontitis (AP) subjects.

    Journal: Clinical and Experimental Immunology

    Article Title: Chemokines in human periodontal disease tissues

    doi: 10.1046/j.1365-2249.2001.01511.x

    Figure Lengend Snippet: The mean (± standard error of the mean) percentage IP-10+ (▪), RANTES+ ( ), MCP-1+ ( ) and MIP-1alpha+ (□) cells in the inflammatory infiltrates of gingival lesions (graded on the size of infiltrate as small in group 1, intermediate in group 2 and extensive in group 3) from periodontitis (AP) subjects.

    Article Snippet: Although the percent MCP-1+ and RANTES+ cells appeared to decrease with increasing inflammation in periodontitis and healthy/gingivitis tissues, respectively, these differences were not significant.

    Techniques:

    Summary diagram . TLR4 or TNFα receptor stimulation increases the activity of NF-κB and the expression of its target genes NR4A1–3 (NR4A), MCP-1, MIP-3α, and Relb. Loss of NR4A potentiates this induction of MCP-1, while reducing the induction of Relb and MIP-3α. Furthermore using mouse embryonic cells, we identify MIP-3α is a novel Relb target gene. Therefore, NR4A receptors act as repressors of inflammatory-NF-κB-driven MCP-1 (indicated by a red T bar) and enhancers of NF-κB-driven Relb and subsequently MIP-3α. Using a NR4A2 DNA-binding mutant reveals that NR4A2 does not require the ability to bind DNA in order to enhance/repress target genes simultaneously.

    Journal: Frontiers in Immunology

    Article Title: NR4A Receptors Differentially Regulate NF-κB Signaling in Myeloid Cells

    doi: 10.3389/fimmu.2017.00007

    Figure Lengend Snippet: Summary diagram . TLR4 or TNFα receptor stimulation increases the activity of NF-κB and the expression of its target genes NR4A1–3 (NR4A), MCP-1, MIP-3α, and Relb. Loss of NR4A potentiates this induction of MCP-1, while reducing the induction of Relb and MIP-3α. Furthermore using mouse embryonic cells, we identify MIP-3α is a novel Relb target gene. Therefore, NR4A receptors act as repressors of inflammatory-NF-κB-driven MCP-1 (indicated by a red T bar) and enhancers of NF-κB-driven Relb and subsequently MIP-3α. Using a NR4A2 DNA-binding mutant reveals that NR4A2 does not require the ability to bind DNA in order to enhance/repress target genes simultaneously.

    Article Snippet: Several studies have shown in both human and mouse macrophages, using NR4A1–3 overexpression and/or knockout cells, that NR4A receptors regulate expression of several downstream NF-κB target genes including IKKi, TNFα, MCP-1, iNos, IP-10, and IL-12 ( , , – ).

    Techniques: Activity Assay, Expressing, Activated Clotting Time Assay, Binding Assay, Mutagenesis

    NR4A2 and 3 negatively regulate TLR4 driven MCP-1 in human and murine myeloid cells . (A,B) Undifferentiated THP-1 cells transduced with shRNA directed against scrambled non-target control, NR4A2, or NR4A3 were treated with 1 µg/ml lipopolysaccharide (LPS) for 18 h (B) and 24 h (A) , followed by media collection and RNA isolation, respectively. (C) Undifferentiated THP-1 cells were pretreated with 10 µM NF-κB inhibitor (NF-κBi) for 1 h followed by the addition of 1 µg/ml LPS for 24 h, followed by media collection. (D) Murine raw mac 264.7 cells transduced with shRNA directed against scrambled non-target control and NR4A2 were exposed to LPS for 1 h. (E) Murine raw mac 264.7 cells transduced with shRNA directed against scrambled non-target control and NR4A2 were exposed to 1 µg/ml LPS for 2 h. (F) Murine raw mac 264.7 cells were pretreated with 10 µM NF-κB inhibitor (NF-κBi) for 1 h followed by exposure to 1 µg/ml LPS for 2 h. Analysis: ELISA analysis was performed for MCP-1 protein detection on media collected at times indicated (A,C) . RNA was isolated and RT-PCR was performed at indicated times to assess levels of MCP-1 NR4A2 and control gene GAPDH (B,D–F) . Un, untreated control. Data are expressed as fold over untreated control (FOC) or pg/ml ± SEM for n = minimum of three individual experiments. * p

    Journal: Frontiers in Immunology

    Article Title: NR4A Receptors Differentially Regulate NF-κB Signaling in Myeloid Cells

    doi: 10.3389/fimmu.2017.00007

    Figure Lengend Snippet: NR4A2 and 3 negatively regulate TLR4 driven MCP-1 in human and murine myeloid cells . (A,B) Undifferentiated THP-1 cells transduced with shRNA directed against scrambled non-target control, NR4A2, or NR4A3 were treated with 1 µg/ml lipopolysaccharide (LPS) for 18 h (B) and 24 h (A) , followed by media collection and RNA isolation, respectively. (C) Undifferentiated THP-1 cells were pretreated with 10 µM NF-κB inhibitor (NF-κBi) for 1 h followed by the addition of 1 µg/ml LPS for 24 h, followed by media collection. (D) Murine raw mac 264.7 cells transduced with shRNA directed against scrambled non-target control and NR4A2 were exposed to LPS for 1 h. (E) Murine raw mac 264.7 cells transduced with shRNA directed against scrambled non-target control and NR4A2 were exposed to 1 µg/ml LPS for 2 h. (F) Murine raw mac 264.7 cells were pretreated with 10 µM NF-κB inhibitor (NF-κBi) for 1 h followed by exposure to 1 µg/ml LPS for 2 h. Analysis: ELISA analysis was performed for MCP-1 protein detection on media collected at times indicated (A,C) . RNA was isolated and RT-PCR was performed at indicated times to assess levels of MCP-1 NR4A2 and control gene GAPDH (B,D–F) . Un, untreated control. Data are expressed as fold over untreated control (FOC) or pg/ml ± SEM for n = minimum of three individual experiments. * p

    Article Snippet: Several studies have shown in both human and mouse macrophages, using NR4A1–3 overexpression and/or knockout cells, that NR4A receptors regulate expression of several downstream NF-κB target genes including IKKi, TNFα, MCP-1, iNos, IP-10, and IL-12 ( , , – ).

    Techniques: Transduction, shRNA, Isolation, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction

    Quantitative chemokine mRNA expression in the CNS. The relative amount of mRNA transcripts for (A) MCP-1 and (B) MIP-1α was analyzed from spinal cord tissue of MBP-EAE (MBP), S100β-EAE (S100), and control animals (control) by dot blot hybridization at days 2 and 4 after transfer. Note the massive up-regulation of MCP-1 and MIP-1α mRNA in the CNS of T MBP cell–treated, but not in the T S100β cell–treated, animals. Amplification of β-tubulin and analysis on ethidium bromide–stained agarose gels demonstrated intact RNA in all samples (not depicted). The data were confirmed in a second set of independently prepared samples.

    Journal: The Journal of Experimental Medicine

    Article Title: The Activation Status of Neuroantigen-specific T Cells in the Target Organ Determines the Clinical Outcome of Autoimmune Encephalomyelitis

    doi: 10.1084/jem.20031064

    Figure Lengend Snippet: Quantitative chemokine mRNA expression in the CNS. The relative amount of mRNA transcripts for (A) MCP-1 and (B) MIP-1α was analyzed from spinal cord tissue of MBP-EAE (MBP), S100β-EAE (S100), and control animals (control) by dot blot hybridization at days 2 and 4 after transfer. Note the massive up-regulation of MCP-1 and MIP-1α mRNA in the CNS of T MBP cell–treated, but not in the T S100β cell–treated, animals. Amplification of β-tubulin and analysis on ethidium bromide–stained agarose gels demonstrated intact RNA in all samples (not depicted). The data were confirmed in a second set of independently prepared samples.

    Article Snippet: In contrast, in rats injected with S100β-specific T cells, there was no corresponding increase in the signals for MCP-1 and MIP-1α, which remained ∼10-fold lower than in animals with MBP-induced EAE ( ).

    Techniques: Expressing, Dot Blot, Hybridization, Amplification, Staining

    MPs are released by stressed adipocytes in a caspase 3 and Rho-associated kinase dependent manner. (A-D) Characterization of adipocyte-derived MPs. (A) Morphology of the differentiated 3T3-L1 adipocytes treated with control, 0.5 mM palmitic acid. (B) The number of Annexin V positive MPs was quantitated by flow cytometry. (C) Dynamic light scattering analysis and Transmission Electron Microscopy of isolated MPs. Isolated MPs were measured by Zetasizer and analyzed using intensity. (D) Western Blot analysis of MPs released by adipocytes. Isolated MPs were fractionated by SDS-PAGE and probed with FABP4, MCP-1, Chemerin, Adiponectin and Perilipin antibody. (E-F) Differentiated adipocytes were incubated with or without palmitic acid in the absence or presence of a selective caspase-3 inhibitor (E), or a range of doses of two different Rho associated kinase inhibitors (Y27632 and fasudil) (F) for up to 12 hrs. Supernatants were then collected and MPs isolated by ultracentrifugation as detailed in methods section. (G) Morphology of isolated mouse primary adipocytes. (H) Number of annexin V positive mouse primary adipocyte-derived MPs assessed by flow cytometry. Values represent mean ± S.D. * P

    Journal: PLoS ONE

    Article Title: Microparticles Release by Adipocytes Act as “Find-Me” Signals to Promote Macrophage Migration

    doi: 10.1371/journal.pone.0123110

    Figure Lengend Snippet: MPs are released by stressed adipocytes in a caspase 3 and Rho-associated kinase dependent manner. (A-D) Characterization of adipocyte-derived MPs. (A) Morphology of the differentiated 3T3-L1 adipocytes treated with control, 0.5 mM palmitic acid. (B) The number of Annexin V positive MPs was quantitated by flow cytometry. (C) Dynamic light scattering analysis and Transmission Electron Microscopy of isolated MPs. Isolated MPs were measured by Zetasizer and analyzed using intensity. (D) Western Blot analysis of MPs released by adipocytes. Isolated MPs were fractionated by SDS-PAGE and probed with FABP4, MCP-1, Chemerin, Adiponectin and Perilipin antibody. (E-F) Differentiated adipocytes were incubated with or without palmitic acid in the absence or presence of a selective caspase-3 inhibitor (E), or a range of doses of two different Rho associated kinase inhibitors (Y27632 and fasudil) (F) for up to 12 hrs. Supernatants were then collected and MPs isolated by ultracentrifugation as detailed in methods section. (G) Morphology of isolated mouse primary adipocytes. (H) Number of annexin V positive mouse primary adipocyte-derived MPs assessed by flow cytometry. Values represent mean ± S.D. * P

    Article Snippet: Macrophages, that infiltrate the adipose tissue of obese mice and humans, are the major source for some of these products, especially of pro-inflammatory cytokines such as TNF-alpha and IL-6 and chemokines such as MCP-1 [ – , ].

    Techniques: Derivative Assay, Flow Cytometry, Cytometry, Transmission Assay, Electron Microscopy, Isolation, Western Blot, SDS Page, Incubation

    Attraction of monocytes and macrophages to supernatants of mature adipocytes exposed to palmitic acid. Migration of (A) RAW264.7 cells or (B) primary mouse macrophages through a transwell (8 μm pore size) to supernatants from untreated (control) or treated differentiated mature adipocytes with 0.5mM palmitic acid. MCP-1 (50 ng/ml) was used as positive control. (C) Adipocytes were treated with 0.5 mM palmitic acid in the absence or presence of 0.025 U/ml apyrase, 0.5 U/ml phospholipase-D or 50 μg/ml control IgG or MCP-1 neutralizing antibody. Macrophages that migrated to the lower chamber were stained with DAPI and the number of cells was counted under fluorescence microscopy. Values represent as mean ± S.D. of representative experiment. * P

    Journal: PLoS ONE

    Article Title: Microparticles Release by Adipocytes Act as “Find-Me” Signals to Promote Macrophage Migration

    doi: 10.1371/journal.pone.0123110

    Figure Lengend Snippet: Attraction of monocytes and macrophages to supernatants of mature adipocytes exposed to palmitic acid. Migration of (A) RAW264.7 cells or (B) primary mouse macrophages through a transwell (8 μm pore size) to supernatants from untreated (control) or treated differentiated mature adipocytes with 0.5mM palmitic acid. MCP-1 (50 ng/ml) was used as positive control. (C) Adipocytes were treated with 0.5 mM palmitic acid in the absence or presence of 0.025 U/ml apyrase, 0.5 U/ml phospholipase-D or 50 μg/ml control IgG or MCP-1 neutralizing antibody. Macrophages that migrated to the lower chamber were stained with DAPI and the number of cells was counted under fluorescence microscopy. Values represent as mean ± S.D. of representative experiment. * P

    Article Snippet: Macrophages, that infiltrate the adipose tissue of obese mice and humans, are the major source for some of these products, especially of pro-inflammatory cytokines such as TNF-alpha and IL-6 and chemokines such as MCP-1 [ – , ].

    Techniques: Migration, Positive Control, Staining, Fluorescence, Microscopy

    Effect of MCP-1 on chemotaxis of CCR2-positive mononuclear cells in viral myocarditis in vitro. Peripheral blood mononuclear cells were separated from mice 7 days post-CVB3 infection. The purified peripheral blood mononuclear cells were analyzed for the

    Journal: Journal of Virology

    Article Title: Coxsackievirus Group B Type 3 Infection Upregulates Expression of Monocyte Chemoattractant Protein 1 in Cardiac Myocytes, Which Leads to Enhanced Migration of Mononuclear Cells in Viral Myocarditis

    doi: 10.1128/JVI.78.22.12548-12556.2004

    Figure Lengend Snippet: Effect of MCP-1 on chemotaxis of CCR2-positive mononuclear cells in viral myocarditis in vitro. Peripheral blood mononuclear cells were separated from mice 7 days post-CVB3 infection. The purified peripheral blood mononuclear cells were analyzed for the

    Article Snippet: The predominant role of MCP-1 in the pathogenesis of viral myocarditis provides a potential target for prevention and therapy of viral myocarditis in the future.

    Techniques: Chemotaxis Assay, In Vitro, Mouse Assay, Infection, Purification

    Effect of anti-MCP-1 on the infiltration of mononuclear cells into myocardial tissue. Male BALB/c mice were inoculated intraperitoneally with CVB3. The mice were given intracardiac injections of anti-MCP-1 antibody or goat IgG on days 3 and 5 postinfection

    Journal: Journal of Virology

    Article Title: Coxsackievirus Group B Type 3 Infection Upregulates Expression of Monocyte Chemoattractant Protein 1 in Cardiac Myocytes, Which Leads to Enhanced Migration of Mononuclear Cells in Viral Myocarditis

    doi: 10.1128/JVI.78.22.12548-12556.2004

    Figure Lengend Snippet: Effect of anti-MCP-1 on the infiltration of mononuclear cells into myocardial tissue. Male BALB/c mice were inoculated intraperitoneally with CVB3. The mice were given intracardiac injections of anti-MCP-1 antibody or goat IgG on days 3 and 5 postinfection

    Article Snippet: The predominant role of MCP-1 in the pathogenesis of viral myocarditis provides a potential target for prevention and therapy of viral myocarditis in the future.

    Techniques: Mouse Assay

    Expression of MCP-1 by myocardial cells infected with wild-type CVB3 or UV-irradiated CVB3. Cardiac myocytes from neonatal BALB/c mice were incubated with UV-irradiated CVB3 (UV-CVB3, ▵) or wild-type CVB3 (WT-CVB3, □) for various periods

    Journal: Journal of Virology

    Article Title: Coxsackievirus Group B Type 3 Infection Upregulates Expression of Monocyte Chemoattractant Protein 1 in Cardiac Myocytes, Which Leads to Enhanced Migration of Mononuclear Cells in Viral Myocarditis

    doi: 10.1128/JVI.78.22.12548-12556.2004

    Figure Lengend Snippet: Expression of MCP-1 by myocardial cells infected with wild-type CVB3 or UV-irradiated CVB3. Cardiac myocytes from neonatal BALB/c mice were incubated with UV-irradiated CVB3 (UV-CVB3, ▵) or wild-type CVB3 (WT-CVB3, □) for various periods

    Article Snippet: The predominant role of MCP-1 in the pathogenesis of viral myocarditis provides a potential target for prevention and therapy of viral myocarditis in the future.

    Techniques: Expressing, Infection, Irradiation, Mouse Assay, Incubation

    Expression of MCP-1 by cardiac myocytes infected with wild-type CVB3. Cardiac myocytes from neonatal BALB/c mice were incubated with wild-type CVB3 (□) or MEM (▪) for various time periods (A and B) or with different CVB3 doses (C and D).

    Journal: Journal of Virology

    Article Title: Coxsackievirus Group B Type 3 Infection Upregulates Expression of Monocyte Chemoattractant Protein 1 in Cardiac Myocytes, Which Leads to Enhanced Migration of Mononuclear Cells in Viral Myocarditis

    doi: 10.1128/JVI.78.22.12548-12556.2004

    Figure Lengend Snippet: Expression of MCP-1 by cardiac myocytes infected with wild-type CVB3. Cardiac myocytes from neonatal BALB/c mice were incubated with wild-type CVB3 (□) or MEM (▪) for various time periods (A and B) or with different CVB3 doses (C and D).

    Article Snippet: The predominant role of MCP-1 in the pathogenesis of viral myocarditis provides a potential target for prevention and therapy of viral myocarditis in the future.

    Techniques: Expressing, Infection, Mouse Assay, Incubation

    Correlation between expression of MCP-1 and infiltration of mononuclear cells in viral myocarditis. Male BALB/c mice were inoculated intraperitoneally with CVB3 or Eagle's medium. At 0, 4, 7, and 9 days postinfection, mice were exsanguinated under ether

    Journal: Journal of Virology

    Article Title: Coxsackievirus Group B Type 3 Infection Upregulates Expression of Monocyte Chemoattractant Protein 1 in Cardiac Myocytes, Which Leads to Enhanced Migration of Mononuclear Cells in Viral Myocarditis

    doi: 10.1128/JVI.78.22.12548-12556.2004

    Figure Lengend Snippet: Correlation between expression of MCP-1 and infiltration of mononuclear cells in viral myocarditis. Male BALB/c mice were inoculated intraperitoneally with CVB3 or Eagle's medium. At 0, 4, 7, and 9 days postinfection, mice were exsanguinated under ether

    Article Snippet: The predominant role of MCP-1 in the pathogenesis of viral myocarditis provides a potential target for prevention and therapy of viral myocarditis in the future.

    Techniques: Expressing, Mouse Assay

    Effects of MCP-1 expression induced by CVB3 infection on chemotaxis of mononuclear cells. Cardiac myocytes were incubated for 6 h in the absence or presence of CVB3. The culture medium were collected (conditioned medium) from cultures of CVB3-infected

    Journal: Journal of Virology

    Article Title: Coxsackievirus Group B Type 3 Infection Upregulates Expression of Monocyte Chemoattractant Protein 1 in Cardiac Myocytes, Which Leads to Enhanced Migration of Mononuclear Cells in Viral Myocarditis

    doi: 10.1128/JVI.78.22.12548-12556.2004

    Figure Lengend Snippet: Effects of MCP-1 expression induced by CVB3 infection on chemotaxis of mononuclear cells. Cardiac myocytes were incubated for 6 h in the absence or presence of CVB3. The culture medium were collected (conditioned medium) from cultures of CVB3-infected

    Article Snippet: The predominant role of MCP-1 in the pathogenesis of viral myocarditis provides a potential target for prevention and therapy of viral myocarditis in the future.

    Techniques: Expressing, Infection, Chemotaxis Assay, Incubation

    MCP-1 and survival

    Journal: Critical care medicine

    Article Title: Streptococcus pneumoniae and Pseudomonas aeruginosa pneumonia induce distinct host responses

    doi: 10.1097/CCM.0b013e3181b4a76b

    Figure Lengend Snippet: MCP-1 and survival

    Article Snippet: Survival studies were also done on MCP-1-/- mice, based upon the cytokine’s significance in both hierarchical clustering and PCA studies.

    Techniques:

    The effect of STAMP2 gene overexpression on inflammatory factors expression in WAT and BAT of ApoE −/− /LDLR −/− mice. A: Immunohistochemical staining for adipose tissue IL-6, TNF-α, MCP-1 and IL-10 (brown staining considered positive staining; scale bar: 50 µm). B: Semiquantification of IL-6, TNF-α, MCP-1 and IL-10 immunohistochemical staining. Data are mean ± SEM (three sections per mice; n = 4 per group). * P

    Journal: PLoS ONE

    Article Title: Overexpressing STAMP2 Improves Insulin Resistance in Diabetic ApoE−/−/LDLR−/− Mice via Macrophage Polarization Shift in Adipose Tissues

    doi: 10.1371/journal.pone.0078903

    Figure Lengend Snippet: The effect of STAMP2 gene overexpression on inflammatory factors expression in WAT and BAT of ApoE −/− /LDLR −/− mice. A: Immunohistochemical staining for adipose tissue IL-6, TNF-α, MCP-1 and IL-10 (brown staining considered positive staining; scale bar: 50 µm). B: Semiquantification of IL-6, TNF-α, MCP-1 and IL-10 immunohistochemical staining. Data are mean ± SEM (three sections per mice; n = 4 per group). * P

    Article Snippet: The sections were incubated with primary rabbit polyclonal anti–STAMP2 (Proteintech Group Inc., Chicago, IL), anti–CD11c (BlueGene, Shanghai, China), anti-CD206 (BlueGene, Shanghai, China), anti-IL-6, anti-TNF-α , anti-MCP-1, anti-IL-10 and primary rat polyclonal anti-F4/80 antibodies (Abcam, Cambridge, MA, USA) overnight and then with a matching biotinylated secondary antibody for 30 min at 37°C.

    Techniques: Over Expression, Expressing, Mouse Assay, Immunohistochemistry, Staining

    A physiologically relevant dose of the formulation can inhibit MCP-1 induced migration of human monocytes. Cellular migration was assessed using THP-1 monocytes that were treated with vehicle (vehicle control) or treated with MCP-1 (20 ng/ml) alone or with MCP-1 (20 ng/ml) in the presence the complete formulation (+PS) or with MCP-1 (20ng/ml) in the presence the formulation lacking PS (-PS) for 3 hours. Monocyte migration is expressed as a fold-change compared to the proportion of cells that moved from the apical compartment into the basolateral compartment in response to MCP-1 alone that has been arbitrarily set as 1. The data are presented as the mean ± SEM from four independent experiments. Statistical analysis was performed using a one-way ANOVA with Dunnett T3 post-hoc analysis where ** p

    Journal: PLoS ONE

    Article Title: A Unique Combination of Nutritionally Active Ingredients Can Prevent Several Key Processes Associated with Atherosclerosis In Vitro

    doi: 10.1371/journal.pone.0151057

    Figure Lengend Snippet: A physiologically relevant dose of the formulation can inhibit MCP-1 induced migration of human monocytes. Cellular migration was assessed using THP-1 monocytes that were treated with vehicle (vehicle control) or treated with MCP-1 (20 ng/ml) alone or with MCP-1 (20 ng/ml) in the presence the complete formulation (+PS) or with MCP-1 (20ng/ml) in the presence the formulation lacking PS (-PS) for 3 hours. Monocyte migration is expressed as a fold-change compared to the proportion of cells that moved from the apical compartment into the basolateral compartment in response to MCP-1 alone that has been arbitrarily set as 1. The data are presented as the mean ± SEM from four independent experiments. Statistical analysis was performed using a one-way ANOVA with Dunnett T3 post-hoc analysis where ** p

    Article Snippet: The formulation inhibits the migration of human monocytes towards MCP-1 The recruitment of monocytes to the activated endothelium in response to chemokines such as MCP-1 is a critical early step in the development of atherosclerosis [ ] and these actions were confirmed in our system as a significant 6.84-fold induction (p < 0.001) in monocyte recruitment in cells treated with MCP-1 alone was observed when compared to the vehicle control ( ).

    Techniques: Migration

    Major bioactive components associated with the complete formulation can inhibit MCP-1 induced migration of human monocytes. Cellular migration was assessed using THP-1 monocytes that were treated with either vehicle (vehicle control) or with MCP-1 (20 ng/ml) or with MCP-1 (20 ng/ml) in the presence of EPA (30 μg/ml), DHA (19.6 μg/ml), (+)-catechin (1.45 μg/ml), stigmasterol (SS, 10 μg/ml), campsterol (CS, 13.9 μg/ml) or β-sitosterol (β-SS, 27.2 μg/ml) for 3 hours. Monocyte migration is expressed as a fold-change compared to the proportion of cells that moved from the apical compartment into the basolateral compartment in response to MCP-1 alone that has been arbitrarily set as 1. The data are presented as the mean ± SEM from four independent experiments. Statistical analysis was performed using a one-way ANOVA with Dunnett post-hoc analysis on Log transformed data where * p

    Journal: PLoS ONE

    Article Title: A Unique Combination of Nutritionally Active Ingredients Can Prevent Several Key Processes Associated with Atherosclerosis In Vitro

    doi: 10.1371/journal.pone.0151057

    Figure Lengend Snippet: Major bioactive components associated with the complete formulation can inhibit MCP-1 induced migration of human monocytes. Cellular migration was assessed using THP-1 monocytes that were treated with either vehicle (vehicle control) or with MCP-1 (20 ng/ml) or with MCP-1 (20 ng/ml) in the presence of EPA (30 μg/ml), DHA (19.6 μg/ml), (+)-catechin (1.45 μg/ml), stigmasterol (SS, 10 μg/ml), campsterol (CS, 13.9 μg/ml) or β-sitosterol (β-SS, 27.2 μg/ml) for 3 hours. Monocyte migration is expressed as a fold-change compared to the proportion of cells that moved from the apical compartment into the basolateral compartment in response to MCP-1 alone that has been arbitrarily set as 1. The data are presented as the mean ± SEM from four independent experiments. Statistical analysis was performed using a one-way ANOVA with Dunnett post-hoc analysis on Log transformed data where * p

    Article Snippet: The formulation inhibits the migration of human monocytes towards MCP-1 The recruitment of monocytes to the activated endothelium in response to chemokines such as MCP-1 is a critical early step in the development of atherosclerosis [ ] and these actions were confirmed in our system as a significant 6.84-fold induction (p < 0.001) in monocyte recruitment in cells treated with MCP-1 alone was observed when compared to the vehicle control ( ).

    Techniques: Migration, Transformation Assay

    Physiologically relevant doses of the formulation can inhibit the IFN-γ induced expression of MCP-1 and ICAM-1 in human macrophages. Gene transcript levels of MCP-1 (a and c) and ICAM-1 (b, d and e) were assessed in PMA differentiated THP-1 macrophages (a, b, c and d) or HMDM (e) that were either treated with vehicle (vehicle control) or with IFN-γ (250 U/ml) or IFN-γ (250 U/ml) in the presence of the complete formulation (+PS; a, b and e) or with IFN-γ (250 U/ml) in the presence of the formulation lacking PS (-PS; c, d and e) for 3 hours. Gene transcript levels were calculated using the comparative Ct method and normalised to GAPDH levels with values from vehicle treated cells given an arbitrary value of 1. The data are presented as the mean ± SEM from three (a, b, c and d) or four (e) independent experiments. Statistical analysis was performed using a one-way ANOVA with Dunnett T3 post-hoc analysis on log-transformed data where * p

    Journal: PLoS ONE

    Article Title: A Unique Combination of Nutritionally Active Ingredients Can Prevent Several Key Processes Associated with Atherosclerosis In Vitro

    doi: 10.1371/journal.pone.0151057

    Figure Lengend Snippet: Physiologically relevant doses of the formulation can inhibit the IFN-γ induced expression of MCP-1 and ICAM-1 in human macrophages. Gene transcript levels of MCP-1 (a and c) and ICAM-1 (b, d and e) were assessed in PMA differentiated THP-1 macrophages (a, b, c and d) or HMDM (e) that were either treated with vehicle (vehicle control) or with IFN-γ (250 U/ml) or IFN-γ (250 U/ml) in the presence of the complete formulation (+PS; a, b and e) or with IFN-γ (250 U/ml) in the presence of the formulation lacking PS (-PS; c, d and e) for 3 hours. Gene transcript levels were calculated using the comparative Ct method and normalised to GAPDH levels with values from vehicle treated cells given an arbitrary value of 1. The data are presented as the mean ± SEM from three (a, b, c and d) or four (e) independent experiments. Statistical analysis was performed using a one-way ANOVA with Dunnett T3 post-hoc analysis on log-transformed data where * p

    Article Snippet: The formulation inhibits the migration of human monocytes towards MCP-1 The recruitment of monocytes to the activated endothelium in response to chemokines such as MCP-1 is a critical early step in the development of atherosclerosis [ ] and these actions were confirmed in our system as a significant 6.84-fold induction (p < 0.001) in monocyte recruitment in cells treated with MCP-1 alone was observed when compared to the vehicle control ( ).

    Techniques: Expressing, Transformation Assay

    Delivery of CCL2-neutralizing antibodies did not significantly affect macrophage infiltration or tumor angiogenesis. (A) Primary tumor tissues were co-immunofluorescent stained for CK5 (green) and F4/80 (red). Secondary antibody controls overlaid with 4′,6-diamidino-2-phenylindole are shown. (B) Primary tumor tissues were immunostained for vWF8. Magnified insert shows positive staining. Staining was quantified by Image J. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Journal: Translational Oncology

    Article Title: Continuous Delivery of Neutralizing Antibodies Elevate CCL2 Levels in Mice Bearing MCF10CA1d Breast Tumor Xenografts

    doi: 10.1016/j.tranon.2017.06.009

    Figure Lengend Snippet: Delivery of CCL2-neutralizing antibodies did not significantly affect macrophage infiltration or tumor angiogenesis. (A) Primary tumor tissues were co-immunofluorescent stained for CK5 (green) and F4/80 (red). Secondary antibody controls overlaid with 4′,6-diamidino-2-phenylindole are shown. (B) Primary tumor tissues were immunostained for vWF8. Magnified insert shows positive staining. Staining was quantified by Image J. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Article Snippet: ELISA of CCL2 Antibody Levels To prepare plates for ELISA analysis of CCL2 antibodies, 96-well high-protein binding plates were incubated with 100 μl/well with 10 ng/ml recombinant human CCL2 (Peprotech, #300-04) diluted in PBS overnight.

    Techniques: Staining, Two Tailed Test

    CCL2-neutralizing antibodies do not significantly affect progression of MCF10A1D breast tumor xenografts. IgG control– or CCL2 antibody–treated mice were analyzed for (A) tumor growth over time; (B) tumor mass at 28 days; (C) malignancy as assessed by H E stain of primary tumor tissues; and (D) lung metastasis, which was assessed by lung whole mount staining. (E) Lung metastasis was validated by H E stain. Representative metastatic lesion is circled. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Journal: Translational Oncology

    Article Title: Continuous Delivery of Neutralizing Antibodies Elevate CCL2 Levels in Mice Bearing MCF10CA1d Breast Tumor Xenografts

    doi: 10.1016/j.tranon.2017.06.009

    Figure Lengend Snippet: CCL2-neutralizing antibodies do not significantly affect progression of MCF10A1D breast tumor xenografts. IgG control– or CCL2 antibody–treated mice were analyzed for (A) tumor growth over time; (B) tumor mass at 28 days; (C) malignancy as assessed by H E stain of primary tumor tissues; and (D) lung metastasis, which was assessed by lung whole mount staining. (E) Lung metastasis was validated by H E stain. Representative metastatic lesion is circled. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Article Snippet: ELISA of CCL2 Antibody Levels To prepare plates for ELISA analysis of CCL2 antibodies, 96-well high-protein binding plates were incubated with 100 μl/well with 10 ng/ml recombinant human CCL2 (Peprotech, #300-04) diluted in PBS overnight.

    Techniques: Mouse Assay, Staining, Two Tailed Test

    Neutralizing antibodies inhibit THP1 cell migration induced by recombinant human CCL2 protein. (A) THP1 cells were stimulated with increasing concentrations of human (hCCL2) or mouse (mCCL2) recombinant CCL2 protein and analyzed for Transwell migration after 2 hours. Representative images of cells migrated into lower chamber are shown. (B) THP1 cells were stimulated with 10 ng/ml hCCL2 or mCCL2 in the presence or absence of increasing doses of CCL2-neutralizing antibodies or control IgG and analyzed for Transwell migration. Representative images of cells treated with CCL2 and 1 μg/ml control IgG or CCL2-neutralizing antibodies. Statistical analysis was performed using one-way ANOVA test with Bonferonni post hoc comparison. Statistical significance was determined by * P

    Journal: Translational Oncology

    Article Title: Continuous Delivery of Neutralizing Antibodies Elevate CCL2 Levels in Mice Bearing MCF10CA1d Breast Tumor Xenografts

    doi: 10.1016/j.tranon.2017.06.009

    Figure Lengend Snippet: Neutralizing antibodies inhibit THP1 cell migration induced by recombinant human CCL2 protein. (A) THP1 cells were stimulated with increasing concentrations of human (hCCL2) or mouse (mCCL2) recombinant CCL2 protein and analyzed for Transwell migration after 2 hours. Representative images of cells migrated into lower chamber are shown. (B) THP1 cells were stimulated with 10 ng/ml hCCL2 or mCCL2 in the presence or absence of increasing doses of CCL2-neutralizing antibodies or control IgG and analyzed for Transwell migration. Representative images of cells treated with CCL2 and 1 μg/ml control IgG or CCL2-neutralizing antibodies. Statistical analysis was performed using one-way ANOVA test with Bonferonni post hoc comparison. Statistical significance was determined by * P

    Article Snippet: ELISA of CCL2 Antibody Levels To prepare plates for ELISA analysis of CCL2 antibodies, 96-well high-protein binding plates were incubated with 100 μl/well with 10 ng/ml recombinant human CCL2 (Peprotech, #300-04) diluted in PBS overnight.

    Techniques: Migration, Recombinant

    Delivery of CCL2-neutralizing antibodies to tumor-bearing mice increased the levels of human CCL2 in blood and in primary tumor tissues. Tumor-bearing mice were treated with control IgG or CCL2-neutralizing antibodies. Four weeks posttransplantation, ELISAs were performed to measure the levels of (A) human CCL2 in blood, (B) human CCL2 in tumor interstitial fluid, and (C) murine CCL2 in blood. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Journal: Translational Oncology

    Article Title: Continuous Delivery of Neutralizing Antibodies Elevate CCL2 Levels in Mice Bearing MCF10CA1d Breast Tumor Xenografts

    doi: 10.1016/j.tranon.2017.06.009

    Figure Lengend Snippet: Delivery of CCL2-neutralizing antibodies to tumor-bearing mice increased the levels of human CCL2 in blood and in primary tumor tissues. Tumor-bearing mice were treated with control IgG or CCL2-neutralizing antibodies. Four weeks posttransplantation, ELISAs were performed to measure the levels of (A) human CCL2 in blood, (B) human CCL2 in tumor interstitial fluid, and (C) murine CCL2 in blood. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Article Snippet: ELISA of CCL2 Antibody Levels To prepare plates for ELISA analysis of CCL2 antibodies, 96-well high-protein binding plates were incubated with 100 μl/well with 10 ng/ml recombinant human CCL2 (Peprotech, #300-04) diluted in PBS overnight.

    Techniques: Mouse Assay, Two Tailed Test

    CCL2 levels in cultured cells are dependent on the presence of neutralizing antibodies. Cultured MCF10CA1d breast cancer cells or hCAF-2300 fibroblasts were treated with control IgG or CCL2-neutralizing antibodies for 24 hours. (A) Medium was analyzed for CCL2 expression by ELISA. (B). Cells were washed to remove antibodies and incubated in serum free condition medium for an additional 24 hours. CCL2 levels in conditioned medium were analyzed by ELISA. Statistical analysis was performed using one-way ANOVA test with Bonferonni post hoc comparison. Statistical significance was determined by P

    Journal: Translational Oncology

    Article Title: Continuous Delivery of Neutralizing Antibodies Elevate CCL2 Levels in Mice Bearing MCF10CA1d Breast Tumor Xenografts

    doi: 10.1016/j.tranon.2017.06.009

    Figure Lengend Snippet: CCL2 levels in cultured cells are dependent on the presence of neutralizing antibodies. Cultured MCF10CA1d breast cancer cells or hCAF-2300 fibroblasts were treated with control IgG or CCL2-neutralizing antibodies for 24 hours. (A) Medium was analyzed for CCL2 expression by ELISA. (B). Cells were washed to remove antibodies and incubated in serum free condition medium for an additional 24 hours. CCL2 levels in conditioned medium were analyzed by ELISA. Statistical analysis was performed using one-way ANOVA test with Bonferonni post hoc comparison. Statistical significance was determined by P

    Article Snippet: ELISA of CCL2 Antibody Levels To prepare plates for ELISA analysis of CCL2 antibodies, 96-well high-protein binding plates were incubated with 100 μl/well with 10 ng/ml recombinant human CCL2 (Peprotech, #300-04) diluted in PBS overnight.

    Techniques: Cell Culture, Expressing, Enzyme-linked Immunosorbent Assay, Incubation

    Osmotic pump delivery of CCL2-neutralizing antibodies in the MCF10CA1d breast tumor xenograft model. (A) Diagram of pump implantation. Mice orthotopically transplanted with fibroblasts and MCF10CA1d breast cancer cells were implanted with osmotic pumps containing CCL2-neutralizing antibodies or isotype control IgG1 for 4 weeks ( N = 5/group). (B) The volume of fluid pumped out was determined by subtracting the residual volume from initial volume after 4 weeks in vivo implantation. Dotted line indicates the expected volume pumped out (155 μl). (C) Measurement CCL2 antibody concentration in blood by ELISA. (D) ELISA analysis of tumor interstitial CCL2 antibody concentration in harvested tumor samples after 4 week s of implantation. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Journal: Translational Oncology

    Article Title: Continuous Delivery of Neutralizing Antibodies Elevate CCL2 Levels in Mice Bearing MCF10CA1d Breast Tumor Xenografts

    doi: 10.1016/j.tranon.2017.06.009

    Figure Lengend Snippet: Osmotic pump delivery of CCL2-neutralizing antibodies in the MCF10CA1d breast tumor xenograft model. (A) Diagram of pump implantation. Mice orthotopically transplanted with fibroblasts and MCF10CA1d breast cancer cells were implanted with osmotic pumps containing CCL2-neutralizing antibodies or isotype control IgG1 for 4 weeks ( N = 5/group). (B) The volume of fluid pumped out was determined by subtracting the residual volume from initial volume after 4 weeks in vivo implantation. Dotted line indicates the expected volume pumped out (155 μl). (C) Measurement CCL2 antibody concentration in blood by ELISA. (D) ELISA analysis of tumor interstitial CCL2 antibody concentration in harvested tumor samples after 4 week s of implantation. Statistical analysis was performed using two-tailed Student’s t test. Statistical significance was determined by P

    Article Snippet: ELISA of CCL2 Antibody Levels To prepare plates for ELISA analysis of CCL2 antibodies, 96-well high-protein binding plates were incubated with 100 μl/well with 10 ng/ml recombinant human CCL2 (Peprotech, #300-04) diluted in PBS overnight.

    Techniques: Mouse Assay, In Vivo, Concentration Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

    Transplantation of rat NPMSCs overexpressing SIRT1 relieves IDD in rats by downregulating the MCP1/CCR2 axis. (A) RT-qPCR and (B) western blot analyses showed increased mRNA and protein expression of MCP1 and CCR2, and reduced mRNA and protein expression of SIRT1 in the IDD group compared with the control group. In contrast, the mRNA and protein expression of MCP1 and CCR2 decreased and mRNA and protein expression of SIRT1 increased in the IDD rats after transplantation of the NPMSCs or SIRT1-overexpressing NPMSCs, n=4. (C) H E staining revealed an amelioration of the disordered intervertebral structure after the transplantation of NPMSCs or SIRT1-overexpressing NPMSCs, n=6. Based on the (D) RT-qPCR and (E) western blot analyses, lower mRNA and protein expression levels of aggrecan, collagen II and Sox-9 were observed in IDD rats compared with in rats of the control group, n=4. (F) TUNEL staining revealed increased cell apoptosis in IDD rats compared with rats in the control group, which was reduced by NPMSC transplantation and even further decreased after the transplantation of SIRT1-overexpressing NPMSCs, n=6. (G) Western blotting showed higher protein expression of Bax and cleaved caspase-3 and lower expression of Bcl-2 in the IDD group compared with in the control group, and these changes were reversed by the transplantation of NPMSCs or SIRT1-overexpressing NPMSCs, n=4. Data in panel F were analyzed using one-way ANOVA; data in panels A, B, C, D, E and G were analyzed using two-way ANOVA. Tukey's multiple comparisons test was applied as the post hoc test. ** P

    Journal: International Journal of Molecular Medicine

    Article Title: Activation of SIRT1 promotes cartilage differentiation and reduces apoptosis of nucleus pulposus mesenchymal stem cells via the MCP1/CCR2 axis in subjects with intervertebral disc degeneration

    doi: 10.3892/ijmm.2020.4668

    Figure Lengend Snippet: Transplantation of rat NPMSCs overexpressing SIRT1 relieves IDD in rats by downregulating the MCP1/CCR2 axis. (A) RT-qPCR and (B) western blot analyses showed increased mRNA and protein expression of MCP1 and CCR2, and reduced mRNA and protein expression of SIRT1 in the IDD group compared with the control group. In contrast, the mRNA and protein expression of MCP1 and CCR2 decreased and mRNA and protein expression of SIRT1 increased in the IDD rats after transplantation of the NPMSCs or SIRT1-overexpressing NPMSCs, n=4. (C) H E staining revealed an amelioration of the disordered intervertebral structure after the transplantation of NPMSCs or SIRT1-overexpressing NPMSCs, n=6. Based on the (D) RT-qPCR and (E) western blot analyses, lower mRNA and protein expression levels of aggrecan, collagen II and Sox-9 were observed in IDD rats compared with in rats of the control group, n=4. (F) TUNEL staining revealed increased cell apoptosis in IDD rats compared with rats in the control group, which was reduced by NPMSC transplantation and even further decreased after the transplantation of SIRT1-overexpressing NPMSCs, n=6. (G) Western blotting showed higher protein expression of Bax and cleaved caspase-3 and lower expression of Bcl-2 in the IDD group compared with in the control group, and these changes were reversed by the transplantation of NPMSCs or SIRT1-overexpressing NPMSCs, n=4. Data in panel F were analyzed using one-way ANOVA; data in panels A, B, C, D, E and G were analyzed using two-way ANOVA. Tukey's multiple comparisons test was applied as the post hoc test. ** P

    Article Snippet: Next, the membranes were blocked with 5% skimmed milk powder for 1 h at room temperature and incubated with primary antibodies (all purchased from Abcam) against MCP1 (1:2,000; cat. no. ab25124), CCR2 (1:1,000; cat. no. ab203128), aggrecan (1:100; cat. no. ab3778), collagen II (1:1,000; cat. no. ab34712), Sry-related HMG box (Sox)-9 (1:1,000; cat. no. ab185966), Bcl-2 (1:1,000; cat. no. ab32124), Bax (1:1,000; cat. no. ab32503), cleaved caspase-3 (1:1,000; cat. no. ab2302), matrix metalloproteinase (MMP)-13 (1:3,000; cat. no. ab39012), tissue inhibitor of metalloproteinase 1 (TIMP-1; 1:1,000; cat. no. ab38978) and SIRT1 (1:2,000; cat. no. ab110304) at 4°C overnight.

    Techniques: Transplantation Assay, Quantitative RT-PCR, Western Blot, Expressing, Staining, TUNEL Assay

    MCP1 reverses the progression of NPMSC cartilage differentiation and the inhibition of NPMSC apoptosis induced by SIRT1 overexpression. (A-D) Compared with the RES group, the RES + MCP1 group displayed decreased mRNA and protein expression levels of aggrecan, collagen II and Sox-9, lower levels of cell apoptosis, decreased protein expression of Bcl-2 and TIMP-1, and increased protein expression of Bax, cleaved caspase-3 and MMP13, based on the results of (A) RT-qPCR, (B and D) western blotting and (C) flow cytometry. Data in panel C were analyzed using a t test; data in panels A, B and D were analyzed using two-way ANOVA. Tukey's multiple comparisons test was applied as the post hoc test. n=3. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Activation of SIRT1 promotes cartilage differentiation and reduces apoptosis of nucleus pulposus mesenchymal stem cells via the MCP1/CCR2 axis in subjects with intervertebral disc degeneration

    doi: 10.3892/ijmm.2020.4668

    Figure Lengend Snippet: MCP1 reverses the progression of NPMSC cartilage differentiation and the inhibition of NPMSC apoptosis induced by SIRT1 overexpression. (A-D) Compared with the RES group, the RES + MCP1 group displayed decreased mRNA and protein expression levels of aggrecan, collagen II and Sox-9, lower levels of cell apoptosis, decreased protein expression of Bcl-2 and TIMP-1, and increased protein expression of Bax, cleaved caspase-3 and MMP13, based on the results of (A) RT-qPCR, (B and D) western blotting and (C) flow cytometry. Data in panel C were analyzed using a t test; data in panels A, B and D were analyzed using two-way ANOVA. Tukey's multiple comparisons test was applied as the post hoc test. n=3. * P

    Article Snippet: Next, the membranes were blocked with 5% skimmed milk powder for 1 h at room temperature and incubated with primary antibodies (all purchased from Abcam) against MCP1 (1:2,000; cat. no. ab25124), CCR2 (1:1,000; cat. no. ab203128), aggrecan (1:100; cat. no. ab3778), collagen II (1:1,000; cat. no. ab34712), Sry-related HMG box (Sox)-9 (1:1,000; cat. no. ab185966), Bcl-2 (1:1,000; cat. no. ab32124), Bax (1:1,000; cat. no. ab32503), cleaved caspase-3 (1:1,000; cat. no. ab2302), matrix metalloproteinase (MMP)-13 (1:3,000; cat. no. ab39012), tissue inhibitor of metalloproteinase 1 (TIMP-1; 1:1,000; cat. no. ab38978) and SIRT1 (1:2,000; cat. no. ab110304) at 4°C overnight.

    Techniques: Inhibition, Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry

    Roles of NPMSCs in IDD, as well as decreased SIRT1 expression and increased expression of genes related to the MCP1/CCR2 axis in LDH were verified. (A) Elongated spindle-shaped passage 2 NPMSCs from patients with LVF and LDH were observed under an optical microscope. (B) Flow cytometry revealed the expression of CD73 and CD90, but not CD34 and CD45, in NPMSCs from the blank group and the LDH group. (C) Alizarin red, (D) oil red O and (E) alcian blue staining revealed the potential of NPMSCs in the blank group and the LDH group to undergo adipogenic, osteoplastic and chondrogenic differentiation. (F) According to the results of the MTT assay, NPMSCs in the LDH group proliferated faster than NPMSCs in the blank group. (G) RT-qPCR revealed lower expression of Oct4 and Nanog mRNA in NPMSCs from the LDH group compared with in the blank group. (H and I) Compared with the blank group, the LDH group displayed lower mRNA and protein expression of SIRT1 and higher mRNA and protein expression of MCP1 and CCR2, based on the results of RT-qPCR and western blot analyses. Two-way ANOVA was used to analyze data in panels F-I. Tukey's multiple comparisons test was applied as the post hoc test. n=3. ** P

    Journal: International Journal of Molecular Medicine

    Article Title: Activation of SIRT1 promotes cartilage differentiation and reduces apoptosis of nucleus pulposus mesenchymal stem cells via the MCP1/CCR2 axis in subjects with intervertebral disc degeneration

    doi: 10.3892/ijmm.2020.4668

    Figure Lengend Snippet: Roles of NPMSCs in IDD, as well as decreased SIRT1 expression and increased expression of genes related to the MCP1/CCR2 axis in LDH were verified. (A) Elongated spindle-shaped passage 2 NPMSCs from patients with LVF and LDH were observed under an optical microscope. (B) Flow cytometry revealed the expression of CD73 and CD90, but not CD34 and CD45, in NPMSCs from the blank group and the LDH group. (C) Alizarin red, (D) oil red O and (E) alcian blue staining revealed the potential of NPMSCs in the blank group and the LDH group to undergo adipogenic, osteoplastic and chondrogenic differentiation. (F) According to the results of the MTT assay, NPMSCs in the LDH group proliferated faster than NPMSCs in the blank group. (G) RT-qPCR revealed lower expression of Oct4 and Nanog mRNA in NPMSCs from the LDH group compared with in the blank group. (H and I) Compared with the blank group, the LDH group displayed lower mRNA and protein expression of SIRT1 and higher mRNA and protein expression of MCP1 and CCR2, based on the results of RT-qPCR and western blot analyses. Two-way ANOVA was used to analyze data in panels F-I. Tukey's multiple comparisons test was applied as the post hoc test. n=3. ** P

    Article Snippet: Next, the membranes were blocked with 5% skimmed milk powder for 1 h at room temperature and incubated with primary antibodies (all purchased from Abcam) against MCP1 (1:2,000; cat. no. ab25124), CCR2 (1:1,000; cat. no. ab203128), aggrecan (1:100; cat. no. ab3778), collagen II (1:1,000; cat. no. ab34712), Sry-related HMG box (Sox)-9 (1:1,000; cat. no. ab185966), Bcl-2 (1:1,000; cat. no. ab32124), Bax (1:1,000; cat. no. ab32503), cleaved caspase-3 (1:1,000; cat. no. ab2302), matrix metalloproteinase (MMP)-13 (1:3,000; cat. no. ab39012), tissue inhibitor of metalloproteinase 1 (TIMP-1; 1:1,000; cat. no. ab38978) and SIRT1 (1:2,000; cat. no. ab110304) at 4°C overnight.

    Techniques: Expressing, Microscopy, Flow Cytometry, Staining, MTT Assay, Quantitative RT-PCR, Western Blot

    Overexpression of SIRT1 and downregulation of the MCP1/CCR2 axis increases NPMSC cartilage differentiation and inhibits NPMSC apoptosis. (A-D) According to RT-qPCR and western blot analyses, the RES group displayed increased mRNA and protein expression of SIRT1, the MCP1 group exhibited increased mRNA and protein expression of MCP1 and CCR2, and the RS102895 group presented decreased mRNA and protein expression of CCR2 compared with the LDH group. (E and F) Compared with the LDH group, the RES and RS102895 groups displayed increased mRNA and protein expression of aggrecan, collagen II and Sox-9, and the MCP1 group exhibited decreased mRNA and protein expression of aggrecan, collagen II and Sox-9, as shown by the results of RT-qPCR and western blot analyses. (G) According to the flow cytometry results, apoptosis was decreased in the RES and RS102895 groups but increased in the MCP1 group compared with the LDH group. (H) Western blot analysis showed decreased expression levels of Bax, cleaved caspase-3 and TIMP-1 and increased levels of Bcl-2 and MMP13 in the RES and RS102895 groups compared with the LDH group; the opposite results were observed in the MCP1 group. Data in panels A, C, D and E were analyzed using t tests, data in panels A, B, E, F and H were analyzed using two-way ANOVA, and data in panel G were analyzed using one-way ANOVA. Tukey's multiple comparisons test was applied as the post hoc test. n=3. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Activation of SIRT1 promotes cartilage differentiation and reduces apoptosis of nucleus pulposus mesenchymal stem cells via the MCP1/CCR2 axis in subjects with intervertebral disc degeneration

    doi: 10.3892/ijmm.2020.4668

    Figure Lengend Snippet: Overexpression of SIRT1 and downregulation of the MCP1/CCR2 axis increases NPMSC cartilage differentiation and inhibits NPMSC apoptosis. (A-D) According to RT-qPCR and western blot analyses, the RES group displayed increased mRNA and protein expression of SIRT1, the MCP1 group exhibited increased mRNA and protein expression of MCP1 and CCR2, and the RS102895 group presented decreased mRNA and protein expression of CCR2 compared with the LDH group. (E and F) Compared with the LDH group, the RES and RS102895 groups displayed increased mRNA and protein expression of aggrecan, collagen II and Sox-9, and the MCP1 group exhibited decreased mRNA and protein expression of aggrecan, collagen II and Sox-9, as shown by the results of RT-qPCR and western blot analyses. (G) According to the flow cytometry results, apoptosis was decreased in the RES and RS102895 groups but increased in the MCP1 group compared with the LDH group. (H) Western blot analysis showed decreased expression levels of Bax, cleaved caspase-3 and TIMP-1 and increased levels of Bcl-2 and MMP13 in the RES and RS102895 groups compared with the LDH group; the opposite results were observed in the MCP1 group. Data in panels A, C, D and E were analyzed using t tests, data in panels A, B, E, F and H were analyzed using two-way ANOVA, and data in panel G were analyzed using one-way ANOVA. Tukey's multiple comparisons test was applied as the post hoc test. n=3. * P

    Article Snippet: Next, the membranes were blocked with 5% skimmed milk powder for 1 h at room temperature and incubated with primary antibodies (all purchased from Abcam) against MCP1 (1:2,000; cat. no. ab25124), CCR2 (1:1,000; cat. no. ab203128), aggrecan (1:100; cat. no. ab3778), collagen II (1:1,000; cat. no. ab34712), Sry-related HMG box (Sox)-9 (1:1,000; cat. no. ab185966), Bcl-2 (1:1,000; cat. no. ab32124), Bax (1:1,000; cat. no. ab32503), cleaved caspase-3 (1:1,000; cat. no. ab2302), matrix metalloproteinase (MMP)-13 (1:3,000; cat. no. ab39012), tissue inhibitor of metalloproteinase 1 (TIMP-1; 1:1,000; cat. no. ab38978) and SIRT1 (1:2,000; cat. no. ab110304) at 4°C overnight.

    Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Expressing, Flow Cytometry

    Overexpression of SIRT1 downregulates the MCP1/CCR2 axis. According to (A) RT-qPCR and (B) western blot analyses, increased mRNA and protein expression of MCP1 and CCR2 were observed in the RES group, and higher mRNA and protein expression of MCP1 and CCR2 were observed in the RES + MCP1 group compared with in the RES group, but the levels were lower than those of the MCP1 group. Two-way ANOVA, followed by Tukey's multiple comparisons test as the post hoc test, were used to determine statistical significance. n=3. ** P

    Journal: International Journal of Molecular Medicine

    Article Title: Activation of SIRT1 promotes cartilage differentiation and reduces apoptosis of nucleus pulposus mesenchymal stem cells via the MCP1/CCR2 axis in subjects with intervertebral disc degeneration

    doi: 10.3892/ijmm.2020.4668

    Figure Lengend Snippet: Overexpression of SIRT1 downregulates the MCP1/CCR2 axis. According to (A) RT-qPCR and (B) western blot analyses, increased mRNA and protein expression of MCP1 and CCR2 were observed in the RES group, and higher mRNA and protein expression of MCP1 and CCR2 were observed in the RES + MCP1 group compared with in the RES group, but the levels were lower than those of the MCP1 group. Two-way ANOVA, followed by Tukey's multiple comparisons test as the post hoc test, were used to determine statistical significance. n=3. ** P

    Article Snippet: Next, the membranes were blocked with 5% skimmed milk powder for 1 h at room temperature and incubated with primary antibodies (all purchased from Abcam) against MCP1 (1:2,000; cat. no. ab25124), CCR2 (1:1,000; cat. no. ab203128), aggrecan (1:100; cat. no. ab3778), collagen II (1:1,000; cat. no. ab34712), Sry-related HMG box (Sox)-9 (1:1,000; cat. no. ab185966), Bcl-2 (1:1,000; cat. no. ab32124), Bax (1:1,000; cat. no. ab32503), cleaved caspase-3 (1:1,000; cat. no. ab2302), matrix metalloproteinase (MMP)-13 (1:3,000; cat. no. ab39012), tissue inhibitor of metalloproteinase 1 (TIMP-1; 1:1,000; cat. no. ab38978) and SIRT1 (1:2,000; cat. no. ab110304) at 4°C overnight.

    Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Expressing

    AS-IV decreased the serum levels of TNF- α , MCP-1, and ICAM-1 in rats with ischemic AKI. Serum levels of TNF- α (a), MCP-1 (b), and ICAM-1 (c) in sham, vehicle-, or AS-IV-pretreated rats at 12 h of reperfusion. Serum levels of TNF- α (d), MCP-1 (e), and ICAM-1 (f) in sham, vehicle-, or AS-IV-pretreated rats at 24 h of reperfusion. Results are expressed as mean ± SD ( n = 8). * P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Preventive Effects of a Natural Anti-Inflammatory Agent, Astragaloside IV, on Ischemic Acute Kidney Injury in Rats

    doi: 10.1155/2013/284025

    Figure Lengend Snippet: AS-IV decreased the serum levels of TNF- α , MCP-1, and ICAM-1 in rats with ischemic AKI. Serum levels of TNF- α (a), MCP-1 (b), and ICAM-1 (c) in sham, vehicle-, or AS-IV-pretreated rats at 12 h of reperfusion. Serum levels of TNF- α (d), MCP-1 (e), and ICAM-1 (f) in sham, vehicle-, or AS-IV-pretreated rats at 24 h of reperfusion. Results are expressed as mean ± SD ( n = 8). * P

    Article Snippet: AS-IV Reduced the Protein Content of MCP-1, ICAM-1, and TNF-α in Rats with Ischemic AKI We observed that the protein content levels of TNF-α , MCP-1 , and ICAM-1 ( ) in renal tissues were elevated at 12 h of reperfusion in rats with ischemic AKI, which were partially restored by AS-IV pretreatment.

    Techniques:

    AS-IV downregulated the mRNA expression of TNF- α , MCP-1, and ICAM-1 in rats with ischemic AKI. Representative real-time PCR of TNF- α (a), MCP-1 (b), and ICAM-1 (c) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 12 h of reperfusion. Representative real-time PCR of TNF- α (d), MCP-1 (e), and ICAM-1 (f) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 24 h of reperfusion. Results are expressed as mean ± SD. * P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Preventive Effects of a Natural Anti-Inflammatory Agent, Astragaloside IV, on Ischemic Acute Kidney Injury in Rats

    doi: 10.1155/2013/284025

    Figure Lengend Snippet: AS-IV downregulated the mRNA expression of TNF- α , MCP-1, and ICAM-1 in rats with ischemic AKI. Representative real-time PCR of TNF- α (a), MCP-1 (b), and ICAM-1 (c) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 12 h of reperfusion. Representative real-time PCR of TNF- α (d), MCP-1 (e), and ICAM-1 (f) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 24 h of reperfusion. Results are expressed as mean ± SD. * P

    Article Snippet: AS-IV Reduced the Protein Content of MCP-1, ICAM-1, and TNF-α in Rats with Ischemic AKI We observed that the protein content levels of TNF-α , MCP-1 , and ICAM-1 ( ) in renal tissues were elevated at 12 h of reperfusion in rats with ischemic AKI, which were partially restored by AS-IV pretreatment.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    AS-IV reduced the protein content of MCP-1, ICAM-1, and TNF- α in rats with ischemic AKI. The protein content of TNF- α (a), MCP-1 (b), and ICAM-1 (c) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 12 h of reperfusion. The protein content of TNF- α (d), MCP-1 (e), and ICAM-1 (f) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 24 h of reperfusion. Results are expressed as mean ± SD ( n = 8). * P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Preventive Effects of a Natural Anti-Inflammatory Agent, Astragaloside IV, on Ischemic Acute Kidney Injury in Rats

    doi: 10.1155/2013/284025

    Figure Lengend Snippet: AS-IV reduced the protein content of MCP-1, ICAM-1, and TNF- α in rats with ischemic AKI. The protein content of TNF- α (a), MCP-1 (b), and ICAM-1 (c) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 12 h of reperfusion. The protein content of TNF- α (d), MCP-1 (e), and ICAM-1 (f) in renal tissues from sham, vehicle-, or AS-IV-pretreated rats at 24 h of reperfusion. Results are expressed as mean ± SD ( n = 8). * P

    Article Snippet: AS-IV Reduced the Protein Content of MCP-1, ICAM-1, and TNF-α in Rats with Ischemic AKI We observed that the protein content levels of TNF-α , MCP-1 , and ICAM-1 ( ) in renal tissues were elevated at 12 h of reperfusion in rats with ischemic AKI, which were partially restored by AS-IV pretreatment.

    Techniques:

    Monocyte chemotactic activity in supernatants of endothelial cells. Monolayers of endothelial cells were stimulated with 100 U/ml recombinant human IL-β, or incubated with 5 × 10 7 strain 42D or with 1 × 10 7 strain CAPD for 60 min at 37°C. After washing and lysostaphin treatment, the endothelial cells were subsequently cultured. At different time points, culture supernatants were harvested and tested for chemotactic activity towards human monocytes. A representative experiment is shown. Data are expressed as the ratio between the number of migrated monocytes towards culture supernatant of infected or IL-1-β-stimulated endothelial cells and the number of migrated monocytes towards culture supernatant of untreated endothelial cells. Chemotaxis was tested in the presence (□) or absence (▪) of neutralizing anti-MCP-1 antibody.

    Journal: Clinical and Experimental Immunology

    Article Title: Infection of human endothelial cells with Staphylococcus aureus induces the production of monocyte chemotactic protein-1 (MCP-1) and monocyte chemotaxis

    doi: 10.1046/j.1365-2249.1999.01002.x

    Figure Lengend Snippet: Monocyte chemotactic activity in supernatants of endothelial cells. Monolayers of endothelial cells were stimulated with 100 U/ml recombinant human IL-β, or incubated with 5 × 10 7 strain 42D or with 1 × 10 7 strain CAPD for 60 min at 37°C. After washing and lysostaphin treatment, the endothelial cells were subsequently cultured. At different time points, culture supernatants were harvested and tested for chemotactic activity towards human monocytes. A representative experiment is shown. Data are expressed as the ratio between the number of migrated monocytes towards culture supernatant of infected or IL-1-β-stimulated endothelial cells and the number of migrated monocytes towards culture supernatant of untreated endothelial cells. Chemotaxis was tested in the presence (□) or absence (▪) of neutralizing anti-MCP-1 antibody.

    Article Snippet: The anti-MCP-1 antibody completely abolished the increased monocyte migration towards recombinant human MCP-1, but had no effect on the fMLP-induced chemotaxis (not shown).

    Techniques: Activity Assay, Recombinant, Incubation, Cell Culture, Infection, Chemotaxis Assay

    Time course of MCP-1 production by endothelial cells. Monolayers of endothelial cells were incubated for 60 min at 37°C with medium alone, with 5 × 10 7 Staphylococcus aureus strain 42D, with 1 × 10 7 S. aureus strain CAPD (a) or with 100 U/ml recombinant human IL-β (b), washed and treated with lysostaphin and cultured for different time periods. The release of MCP-1 into the culture medium was measured and plotted against time of culture. Values represent means ± s.e.m. of three experiments with endothelial cells from different donors.

    Journal: Clinical and Experimental Immunology

    Article Title: Infection of human endothelial cells with Staphylococcus aureus induces the production of monocyte chemotactic protein-1 (MCP-1) and monocyte chemotaxis

    doi: 10.1046/j.1365-2249.1999.01002.x

    Figure Lengend Snippet: Time course of MCP-1 production by endothelial cells. Monolayers of endothelial cells were incubated for 60 min at 37°C with medium alone, with 5 × 10 7 Staphylococcus aureus strain 42D, with 1 × 10 7 S. aureus strain CAPD (a) or with 100 U/ml recombinant human IL-β (b), washed and treated with lysostaphin and cultured for different time periods. The release of MCP-1 into the culture medium was measured and plotted against time of culture. Values represent means ± s.e.m. of three experiments with endothelial cells from different donors.

    Article Snippet: The anti-MCP-1 antibody completely abolished the increased monocyte migration towards recombinant human MCP-1, but had no effect on the fMLP-induced chemotaxis (not shown).

    Techniques: Incubation, Recombinant, Cell Culture

    Lentiviral-mediated shRNA knockdown of endogenous GPSM3 expression in the human monocytic THP-1 cell line disrupts migration toward MCP-1 ( A ) Western blot showing whole lysate (40 μg) immunoreactivity with mouse monoclonal anti-GPSM3 antibody 35.5.1 (ref. 47 ) and with anti-β-actin as a loading control. ( B ) Real-time Transwell migration analysis of indicated calcein-AM stained THP-1 cell lines as shown by change in the ratio of MCP-1-induced to vehicle-induced relative fluorescence units (RFU) over a 30 minute timeframe. ( C ) qRT-PCR data from indicated stable knockdown or scrambled control THP-1 cell line total RNA preparations. There was no significant difference in CCR2 mRNA abundance in the THP-1 cell lines as determined by one-way ANOVA (F = 2.40; p = 0.119). ( D ) Regression analysis-derived rate of THP-1 cell line transmigration over initial migratory period; differences between each cell line was calculated by one-way ANOVA. ( E ) Regression analysis-derived maximum THP-1 cell transmigration over 30 minute timecourese; differences between each cell line were calculated by one-way ANOVA. All data are reported as means with error bars representing S.E.M.

    Journal: Genes and immunity

    Article Title: Genetic variations in GPSM3 associated with protection from rheumatoid arthritis affect its transcript abundance

    doi: 10.1038/gene.2016.3

    Figure Lengend Snippet: Lentiviral-mediated shRNA knockdown of endogenous GPSM3 expression in the human monocytic THP-1 cell line disrupts migration toward MCP-1 ( A ) Western blot showing whole lysate (40 μg) immunoreactivity with mouse monoclonal anti-GPSM3 antibody 35.5.1 (ref. 47 ) and with anti-β-actin as a loading control. ( B ) Real-time Transwell migration analysis of indicated calcein-AM stained THP-1 cell lines as shown by change in the ratio of MCP-1-induced to vehicle-induced relative fluorescence units (RFU) over a 30 minute timeframe. ( C ) qRT-PCR data from indicated stable knockdown or scrambled control THP-1 cell line total RNA preparations. There was no significant difference in CCR2 mRNA abundance in the THP-1 cell lines as determined by one-way ANOVA (F = 2.40; p = 0.119). ( D ) Regression analysis-derived rate of THP-1 cell line transmigration over initial migratory period; differences between each cell line was calculated by one-way ANOVA. ( E ) Regression analysis-derived maximum THP-1 cell transmigration over 30 minute timecourese; differences between each cell line were calculated by one-way ANOVA. All data are reported as means with error bars representing S.E.M.

    Article Snippet: Cells were allowed to migrate toward vehicle or 100 ng/mL MCP-1/CCL2 (R & D Systems, Minneapolis, MN) in the lower chamber.

    Techniques: shRNA, Expressing, Migration, Western Blot, Staining, Fluorescence, Quantitative RT-PCR, Derivative Assay, Transmigration Assay

    IVC levels of TNF-α, IF-γ, MCP-1 and IL-10 in normal, RAS and RAS + MSC pigs ( A – D ). Urine levels of the same cytokines did not differ among the groups ( E – H ). Net release of TNF-α, IF-γ, MCP-1 and IL-10 in

    Journal: Nephrology Dialysis Transplantation

    Article Title: Renal vein cytokine release as an index of renal parenchymal inflammation in chronic experimental renal artery stenosis

    doi: 10.1093/ndt/gft305

    Figure Lengend Snippet: IVC levels of TNF-α, IF-γ, MCP-1 and IL-10 in normal, RAS and RAS + MSC pigs ( A – D ). Urine levels of the same cytokines did not differ among the groups ( E – H ). Net release of TNF-α, IF-γ, MCP-1 and IL-10 in

    Article Snippet: The net release of TNF-α, IF-γ and MCP-1 from the RV was higher in the RAS kidney compared with normal and decreased in MSC-treated pigs (Figure I–K).

    Techniques:

    A direct correlation was found between net renal release of TNF-α, IF-γ and MCP-1 and their tissue expression ( A , C and E , left), but not with their urinary levels ( B , D and F , right). Correlations between net release of

    Journal: Nephrology Dialysis Transplantation

    Article Title: Renal vein cytokine release as an index of renal parenchymal inflammation in chronic experimental renal artery stenosis

    doi: 10.1093/ndt/gft305

    Figure Lengend Snippet: A direct correlation was found between net renal release of TNF-α, IF-γ and MCP-1 and their tissue expression ( A , C and E , left), but not with their urinary levels ( B , D and F , right). Correlations between net release of

    Article Snippet: The net release of TNF-α, IF-γ and MCP-1 from the RV was higher in the RAS kidney compared with normal and decreased in MSC-treated pigs (Figure I–K).

    Techniques: Expressing

    Representative immunoblots ( A ) and renal protein expression ( B ) of TNF-α, IF-γ, MCP-1 and IL-10 in the study groups. *P≤0.05 versus normal, ‡ P

    Journal: Nephrology Dialysis Transplantation

    Article Title: Renal vein cytokine release as an index of renal parenchymal inflammation in chronic experimental renal artery stenosis

    doi: 10.1093/ndt/gft305

    Figure Lengend Snippet: Representative immunoblots ( A ) and renal protein expression ( B ) of TNF-α, IF-γ, MCP-1 and IL-10 in the study groups. *P≤0.05 versus normal, ‡ P

    Article Snippet: The net release of TNF-α, IF-γ and MCP-1 from the RV was higher in the RAS kidney compared with normal and decreased in MSC-treated pigs (Figure I–K).

    Techniques: Western Blot, Expressing

    Emodin reduces E-selectin and MCP-1 expression in the lung tissues in LPS-induced ALI. After 72 h interventions, mice were exsanguinated and their lungs were removed. ( A , C ) qPCR was performed to analyze E-selectin and MCP-1 mRNA expression in the lung tissues; ( B , D ) Western blotting was performed to measure E-selectin and MCP-1 protein expression in the lung tissues. Each bar represents the mean ± SD of 10 mice. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Emodin Ameliorates LPS-Induced Acute Lung Injury, Involving the Inactivation of NF-κB in Mice

    doi: 10.3390/ijms151119355

    Figure Lengend Snippet: Emodin reduces E-selectin and MCP-1 expression in the lung tissues in LPS-induced ALI. After 72 h interventions, mice were exsanguinated and their lungs were removed. ( A , C ) qPCR was performed to analyze E-selectin and MCP-1 mRNA expression in the lung tissues; ( B , D ) Western blotting was performed to measure E-selectin and MCP-1 protein expression in the lung tissues. Each bar represents the mean ± SD of 10 mice. * p

    Article Snippet: After incubation for 1 h in blocking solution at room temperature, the membrane was incubated for 24 h with anti-E-selectin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-MCP-1 (Santa Cruz Biotechnology, Inc.), anti-phospho-NF-κB p65 (Santa Cruz Biotechnology, Inc.), anti-NF-κB p65 (Santa Cruz Biotechnology, Inc.), or anti-β-actin (Santa Cruz Biotechnology, Inc.) at 4 °C, respectively.

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

    Release of cytokines by purified natural killer (NK) cells upon stimulation with selected expansion protocols. Purified NK cells were cultured using different stimulation protocols. On day 6, release of tumor necrosis factor (TNF)-α, IFN-γ, macrophage inflammatory protein (MIP)-1α, monocyte chemoattractant protein (MCP)-1, granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-8, and IL-10 were assayed by cytometric bead array analyses. Graphs show mean values and SDs obtained from four independent donors, except GM-CSF, which was measured for three independent donors. Asterisks indicate significant differences (* p

    Journal: Frontiers in Immunology

    Article Title: A Two-Phase Expansion Protocol Combining Interleukin (IL)-15 and IL-21 Improves Natural Killer Cell Proliferation and Cytotoxicity against Rhabdomyosarcoma

    doi: 10.3389/fimmu.2017.00676

    Figure Lengend Snippet: Release of cytokines by purified natural killer (NK) cells upon stimulation with selected expansion protocols. Purified NK cells were cultured using different stimulation protocols. On day 6, release of tumor necrosis factor (TNF)-α, IFN-γ, macrophage inflammatory protein (MIP)-1α, monocyte chemoattractant protein (MCP)-1, granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-8, and IL-10 were assayed by cytometric bead array analyses. Graphs show mean values and SDs obtained from four independent donors, except GM-CSF, which was measured for three independent donors. Asterisks indicate significant differences (* p

    Article Snippet: Cytometric Bead Array Cytokine secretion was examined by cytometric bead array analyses (CBA) on supernatants of stimulated NK cells on the sixth day of cultivation using BD CBA Flex Sets for IFN-γ, tumor necrosis factor (TNF)-α, macrophage inflammatory protein (MIP)-1α, monocyte chemoattractant protein (MCP)-1, IL-8, IL-10, and granulocyte-macrophage colony-stimulating factor (GM-CSF) (BD Biosciences).

    Techniques: Purification, Cell Culture

    Activation of the FoxO1–KLF2–S1P1 axis by CCR2 signaling through Stat3. (A) Surface expression of S1P1 in the CD69 + Qa-2 − (SP1 + SP2), CD69 − Qa-2 − (SP3), and CD69 − Qa-2 + (SP4) subsets of CD4 SP thymocytes from WT and KO mice. The experiments were repeated three times with six mice for each group. Representative dot plots are shown. (B) Percentage of S1P1 + cells are presented as mean ± SD. (C) Foxo1, Klf2, and S1pr1 mRNA expression in WT and KO CD4 SP thymocytes as determined by qPCR. The experiments were repeated three times with triplicates. Data are present as mean ± SEM. (D) CD69 − Qa-2 + SP4 thymocytes from WT and KO mice were stimulated with CCL2 in the presence or absence of STAT3 inhibitor Stattic. Foxo1, Klf2, and S1pr1 mRNA expression was detected by qPCR. The experiments were repeated three times with triplicates. Data are present as mean ± SEM. (E) Following stimulation with CCL2, WT SP4 thymocytes were examined for surface expression of S1P1. Representative histograms are shown out of three independent experiments. (F,G) Levels of phosphorylated Stat3 were examined by Western blotting (F) or intracellular staining (G) in SP4 thymocytes after stimulation with CCL2. Western blotting was performed at 30 min after stimulation. The experiments were repeated three times with similar results. (H–J) SP4 thymocytes were stimulated with CCL2 in the presence or absence of STAT3 inhibitor Stattic. Surface expression of S1P1 was assayed by flow cytometry (H) . Subcellular localization of FoxO1 was revealed by immunofluorescent staining (I) and nucleus Foxo1 was quantified by mean fluorescent intensity (MFI) (J) . Data are presented as mean ± SD. Two independent experiments were performed with similar results. * p

    Journal: Frontiers in Immunology

    Article Title: CCR2 Signal Facilitates Thymic Egress by Priming Thymocyte Responses to Sphingosine-1-Phosphate

    doi: 10.3389/fimmu.2018.01263

    Figure Lengend Snippet: Activation of the FoxO1–KLF2–S1P1 axis by CCR2 signaling through Stat3. (A) Surface expression of S1P1 in the CD69 + Qa-2 − (SP1 + SP2), CD69 − Qa-2 − (SP3), and CD69 − Qa-2 + (SP4) subsets of CD4 SP thymocytes from WT and KO mice. The experiments were repeated three times with six mice for each group. Representative dot plots are shown. (B) Percentage of S1P1 + cells are presented as mean ± SD. (C) Foxo1, Klf2, and S1pr1 mRNA expression in WT and KO CD4 SP thymocytes as determined by qPCR. The experiments were repeated three times with triplicates. Data are present as mean ± SEM. (D) CD69 − Qa-2 + SP4 thymocytes from WT and KO mice were stimulated with CCL2 in the presence or absence of STAT3 inhibitor Stattic. Foxo1, Klf2, and S1pr1 mRNA expression was detected by qPCR. The experiments were repeated three times with triplicates. Data are present as mean ± SEM. (E) Following stimulation with CCL2, WT SP4 thymocytes were examined for surface expression of S1P1. Representative histograms are shown out of three independent experiments. (F,G) Levels of phosphorylated Stat3 were examined by Western blotting (F) or intracellular staining (G) in SP4 thymocytes after stimulation with CCL2. Western blotting was performed at 30 min after stimulation. The experiments were repeated three times with similar results. (H–J) SP4 thymocytes were stimulated with CCL2 in the presence or absence of STAT3 inhibitor Stattic. Surface expression of S1P1 was assayed by flow cytometry (H) . Subcellular localization of FoxO1 was revealed by immunofluorescent staining (I) and nucleus Foxo1 was quantified by mean fluorescent intensity (MFI) (J) . Data are presented as mean ± SD. Two independent experiments were performed with similar results. * p

    Article Snippet: Recombinant mouse CCL2, CXCL9, and CXCL16 were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Activation Assay, Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Western Blot, Staining, Flow Cytometry, Cytometry

    Restricted expression of CCR2 in CD69 − Qa-2 + mature CD4 single-positive (SP) thymocytes. (A,B) Genes encoding chemoattractant receptors were sorted out from the genes differentially expressed among the four subsets (SP1–SP4) of CD4 SP thymocytes. The expression patterns of these genes are depicted in heatmap (A) or line charts (B) . (C) Expression levels of Cxcr3, Cxcr6, S1pr4 , and Ccr2 were assessed in various subsets of CD4 SP thymocytes using qPCR. The experiments were repeated three times with triplicates for each sample and the data are presented as mean ± SEM. (D) CCR2 expression in CD69 + Qa-2 − (SP1 + SP2), CD69 − Qa-2 − (SP3), and CD69 − Qa-2 + (SP4) subsets of CD4 SP thymocytes as determined by flow cytometry. Representative dot plots are shown out of three independent experiments. (E) Purified CD69 − Qa-2 + SP4 thymocytes were tested for chemotaxis to CCL2 and S1P using transwell migration assay. Data from four independent experiments with duplicates are presented as mean ± SEM. Statistical differences between groups were determined by the Student’s t test. ** p

    Journal: Frontiers in Immunology

    Article Title: CCR2 Signal Facilitates Thymic Egress by Priming Thymocyte Responses to Sphingosine-1-Phosphate

    doi: 10.3389/fimmu.2018.01263

    Figure Lengend Snippet: Restricted expression of CCR2 in CD69 − Qa-2 + mature CD4 single-positive (SP) thymocytes. (A,B) Genes encoding chemoattractant receptors were sorted out from the genes differentially expressed among the four subsets (SP1–SP4) of CD4 SP thymocytes. The expression patterns of these genes are depicted in heatmap (A) or line charts (B) . (C) Expression levels of Cxcr3, Cxcr6, S1pr4 , and Ccr2 were assessed in various subsets of CD4 SP thymocytes using qPCR. The experiments were repeated three times with triplicates for each sample and the data are presented as mean ± SEM. (D) CCR2 expression in CD69 + Qa-2 − (SP1 + SP2), CD69 − Qa-2 − (SP3), and CD69 − Qa-2 + (SP4) subsets of CD4 SP thymocytes as determined by flow cytometry. Representative dot plots are shown out of three independent experiments. (E) Purified CD69 − Qa-2 + SP4 thymocytes were tested for chemotaxis to CCL2 and S1P using transwell migration assay. Data from four independent experiments with duplicates are presented as mean ± SEM. Statistical differences between groups were determined by the Student’s t test. ** p

    Article Snippet: Recombinant mouse CCL2, CXCL9, and CXCL16 were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry, Purification, Chemotaxis Assay, Transwell Migration Assay

    CCR2-mediated chemokinesis and its impact on S1P-induced chemotaxis. (A) CCL2 concentration in the thymic homogenate and serum of wild-type (WT) mice as measured by ELISA. The experiments were repeated three times with duplicates. Data are presented as mean ± SEM. (B) Double-positive thymocytes, and Qa-2 − (SP1–3) and CD69 − Qa-2 + CD4 single-positive (SP) thymocytes were purified from wild-type (WT) and knockout (KO) mice and loaded onto thymic slices. Their migration speed were recorded using two-photon microscopy. The experiments were repeated three times. Each dot represents a single cell (WT SP1–3, n = 171; KO SP1–3, n = 120; WT SP4, n = 193; KO SP4, n = 198). Bars indicate the median. (C) Mean squared displacement (MSD) over a period of 20 min for SP4 thymocytes. Data from more than 100 cells at each time point are presented as mean ± SD. (D) CD69 − Qa-2 + CD4 SP thymocytes were plated in a two-dimensional migration assay. Cell movement under different conditions was tracked using time-lapse imaging. The trajectories of WT (red) and KO (green) CD4 SP4 cells are shown ( Upper ). The displacement of the WT SP4 cells from the original position is depicted at the Bottom . (E) Velocity of cell movement in the two-dimensional migration assay. The experiments were repeated three times. Each dot represents a single cell. Bars indicate the median. * p

    Journal: Frontiers in Immunology

    Article Title: CCR2 Signal Facilitates Thymic Egress by Priming Thymocyte Responses to Sphingosine-1-Phosphate

    doi: 10.3389/fimmu.2018.01263

    Figure Lengend Snippet: CCR2-mediated chemokinesis and its impact on S1P-induced chemotaxis. (A) CCL2 concentration in the thymic homogenate and serum of wild-type (WT) mice as measured by ELISA. The experiments were repeated three times with duplicates. Data are presented as mean ± SEM. (B) Double-positive thymocytes, and Qa-2 − (SP1–3) and CD69 − Qa-2 + CD4 single-positive (SP) thymocytes were purified from wild-type (WT) and knockout (KO) mice and loaded onto thymic slices. Their migration speed were recorded using two-photon microscopy. The experiments were repeated three times. Each dot represents a single cell (WT SP1–3, n = 171; KO SP1–3, n = 120; WT SP4, n = 193; KO SP4, n = 198). Bars indicate the median. (C) Mean squared displacement (MSD) over a period of 20 min for SP4 thymocytes. Data from more than 100 cells at each time point are presented as mean ± SD. (D) CD69 − Qa-2 + CD4 SP thymocytes were plated in a two-dimensional migration assay. Cell movement under different conditions was tracked using time-lapse imaging. The trajectories of WT (red) and KO (green) CD4 SP4 cells are shown ( Upper ). The displacement of the WT SP4 cells from the original position is depicted at the Bottom . (E) Velocity of cell movement in the two-dimensional migration assay. The experiments were repeated three times. Each dot represents a single cell. Bars indicate the median. * p

    Article Snippet: Recombinant mouse CCL2, CXCL9, and CXCL16 were purchased from R & D Systems (Minneapolis, MN, USA).

    Techniques: Chemotaxis Assay, Concentration Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Purification, Knock-Out, Migration, Microscopy, Imaging

    miR-K10a-mediated downregulation of TWEAKR inhibits production of IL-8 and MCP-1. (A) HUVECs were transfected with miR-K10a and treated with TWEAK at 24 hpt. Culture supernatants were harvested at 24 h posttreatment and analyzed for IL-8 (top) and MCP-1

    Journal: Journal of Virology

    Article Title: Regulation of Tumor Necrosis Factor-Like Weak Inducer of Apoptosis Receptor Protein (TWEAKR) Expression by Kaposi's Sarcoma-Associated Herpesvirus MicroRNA Prevents TWEAK-Induced Apoptosis and Inflammatory Cytokine Expression ▿Regulation of Tumor Necrosis Factor-Like Weak Inducer of Apoptosis Receptor Protein (TWEAKR) Expression by Kaposi's Sarcoma-Associated Herpesvirus MicroRNA Prevents TWEAK-Induced Apoptosis and Inflammatory Cytokine Expression ▿ †

    doi: 10.1128/JVI.00884-10

    Figure Lengend Snippet: miR-K10a-mediated downregulation of TWEAKR inhibits production of IL-8 and MCP-1. (A) HUVECs were transfected with miR-K10a and treated with TWEAK at 24 hpt. Culture supernatants were harvested at 24 h posttreatment and analyzed for IL-8 (top) and MCP-1

    Article Snippet: The following primer sets were used: 5′-CTTCAACCTCTTTACCACAAAAGATTC-3′ and 5′-TGCTGGTAGAGTTCGGTGCA-3′ for IFN-α1, 5′-AAATGCCAGCCTGCTGACGAAG-3′ and 5′-AACAACAATCTGAGGTGCCCATGCTAC-3′ for IL-6, 5′-TTGGCAGCCTTCCTGATTTC-3′ and 5′-AACTTCTCCACAACCCTCTG-3′ for IL-8, 5′-CCCCAGTCACCTGCTGTTAT-3′ and 5′-TGGAATCCTGAACCCACTTC-3′ for MCP-1, and 5′-GTGACATTAAGGAGAAGCTGTGCTA-3′ and 5′-CTTCATGATGGGAGTTGAAGGTAGTT-3′ for β-actin.

    Techniques: Transfection