recombinant proteins Search Results


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  • 99
    Thermo Fisher recombinant proteins
    Recombinant Proteins, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1595 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore recombinant proteins
    Recombinant Proteins, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1870 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems il 17a
    Rhodomyrtone inhibits <t>TNF-α+IL-17A-induced</t> inflammatory responses in skin organ cultures. . Bars indicate mean ± SD, n = 5. Statistical significance determined by one-way ANOVA and Dunn’s multiple comparison test, indicated as* p
    Il 17a, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 3062 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher basic fibroblast growth factor
    Rhodomyrtone inhibits <t>TNF-α+IL-17A-induced</t> inflammatory responses in skin organ cultures. . Bars indicate mean ± SD, n = 5. Statistical significance determined by one-way ANOVA and Dunn’s multiple comparison test, indicated as* p
    Basic Fibroblast Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3713 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore recombinant protein
    Rhodomyrtone inhibits <t>TNF-α+IL-17A-induced</t> inflammatory responses in skin organ cultures. . Bars indicate mean ± SD, n = 5. Statistical significance determined by one-way ANOVA and Dunn’s multiple comparison test, indicated as* p
    Recombinant Protein, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher tnf α
    VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) <t>TNF-α,</t> ( b ) IL-6, ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p
    Tnf α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 21879 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems egf
    VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) <t>TNF-α,</t> ( b ) IL-6, ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p
    Egf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 3130 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems netrin 1
    <t>Netrin-1</t> promotes inflammation resolution in diabetic corneal wound healing. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). Mouse cornea was collected and immunofluorescent stained with neutrophil marker Ly6G at 48 h and M2 macrophage markers CD68 (green fluorescence) and CD206 (red fluorescence) at 72 h after epithelial scrape ( A , 3 mice per group). Flow cytometry was performed to quantify the number and percentage of neutrophils (CD45 + CD11b + Ly6G+) ( B , 12 mice per group) at 48-hour post-wounded, M1 macrophages (CD45 + F4/80 + CD86+) and M2 macrophages (CD45 + F4/80 + CD206+) at 72-hour post-wounded. ( C , 12 mice per group at each detection) mRNA expression levels of iNOS and IL-12 (48 h after epithelial scrape), arginase-1 and IL-10 (72 h after epithelial scrape) were analyzed by RT-qPCR from the control and diabetic mouse corneas ( D , 6 mice per group); *p
    Netrin 1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 927 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human tgf β1
    <t>Netrin-1</t> promotes inflammation resolution in diabetic corneal wound healing. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). Mouse cornea was collected and immunofluorescent stained with neutrophil marker Ly6G at 48 h and M2 macrophage markers CD68 (green fluorescence) and CD206 (red fluorescence) at 72 h after epithelial scrape ( A , 3 mice per group). Flow cytometry was performed to quantify the number and percentage of neutrophils (CD45 + CD11b + Ly6G+) ( B , 12 mice per group) at 48-hour post-wounded, M1 macrophages (CD45 + F4/80 + CD86+) and M2 macrophages (CD45 + F4/80 + CD206+) at 72-hour post-wounded. ( C , 12 mice per group at each detection) mRNA expression levels of iNOS and IL-12 (48 h after epithelial scrape), arginase-1 and IL-10 (72 h after epithelial scrape) were analyzed by RT-qPCR from the control and diabetic mouse corneas ( D , 6 mice per group); *p
    Recombinant Human Tgf β1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 2249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems timp 1
    <t>Netrin-1</t> promotes inflammation resolution in diabetic corneal wound healing. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). Mouse cornea was collected and immunofluorescent stained with neutrophil marker Ly6G at 48 h and M2 macrophage markers CD68 (green fluorescence) and CD206 (red fluorescence) at 72 h after epithelial scrape ( A , 3 mice per group). Flow cytometry was performed to quantify the number and percentage of neutrophils (CD45 + CD11b + Ly6G+) ( B , 12 mice per group) at 48-hour post-wounded, M1 macrophages (CD45 + F4/80 + CD86+) and M2 macrophages (CD45 + F4/80 + CD206+) at 72-hour post-wounded. ( C , 12 mice per group at each detection) mRNA expression levels of iNOS and IL-12 (48 h after epithelial scrape), arginase-1 and IL-10 (72 h after epithelial scrape) were analyzed by RT-qPCR from the control and diabetic mouse corneas ( D , 6 mice per group); *p
    Timp 1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1606 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher il 10
    <t>Netrin-1</t> promotes inflammation resolution in diabetic corneal wound healing. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). Mouse cornea was collected and immunofluorescent stained with neutrophil marker Ly6G at 48 h and M2 macrophage markers CD68 (green fluorescence) and CD206 (red fluorescence) at 72 h after epithelial scrape ( A , 3 mice per group). Flow cytometry was performed to quantify the number and percentage of neutrophils (CD45 + CD11b + Ly6G+) ( B , 12 mice per group) at 48-hour post-wounded, M1 macrophages (CD45 + F4/80 + CD86+) and M2 macrophages (CD45 + F4/80 + CD206+) at 72-hour post-wounded. ( C , 12 mice per group at each detection) mRNA expression levels of iNOS and IL-12 (48 h after epithelial scrape), arginase-1 and IL-10 (72 h after epithelial scrape) were analyzed by RT-qPCR from the control and diabetic mouse corneas ( D , 6 mice per group); *p
    Il 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2937 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher human recombinant epidermal growth factor
    <t>Netrin-1</t> promotes inflammation resolution in diabetic corneal wound healing. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). Mouse cornea was collected and immunofluorescent stained with neutrophil marker Ly6G at 48 h and M2 macrophage markers CD68 (green fluorescence) and CD206 (red fluorescence) at 72 h after epithelial scrape ( A , 3 mice per group). Flow cytometry was performed to quantify the number and percentage of neutrophils (CD45 + CD11b + Ly6G+) ( B , 12 mice per group) at 48-hour post-wounded, M1 macrophages (CD45 + F4/80 + CD86+) and M2 macrophages (CD45 + F4/80 + CD206+) at 72-hour post-wounded. ( C , 12 mice per group at each detection) mRNA expression levels of iNOS and IL-12 (48 h after epithelial scrape), arginase-1 and IL-10 (72 h after epithelial scrape) were analyzed by RT-qPCR from the control and diabetic mouse corneas ( D , 6 mice per group); *p
    Human Recombinant Epidermal Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1506 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human tnf α
    The level of <t>TNF-α</t> protein increased in TOLF. A) Fold change of TNF-α protein in the ossified ligamentum flavum of TOLF. Proteins were extracted from the ossified ligamentum flavum (OS-Lig) and normal ligamentum flavum (Lig). The samples were then labeled with iTRAQ following the manufacture’s protocol. *: A star indicates statistical significance compared to control group with p
    Recombinant Human Tnf α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 1509 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher granulocyte macrophage colony stimulating factor
    The level of <t>TNF-α</t> protein increased in TOLF. A) Fold change of TNF-α protein in the ossified ligamentum flavum of TOLF. Proteins were extracted from the ossified ligamentum flavum (OS-Lig) and normal ligamentum flavum (Lig). The samples were then labeled with iTRAQ following the manufacture’s protocol. *: A star indicates statistical significance compared to control group with p
    Granulocyte Macrophage Colony Stimulating Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 390 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human il 6
    3.2. Association of extracellular histones, HMGB1, sTM, and <t>IL-6</t> with ALI
    Human Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 1161 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher interleukin 6
    VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) TNF-α, ( b ) <t>IL-6,</t> ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p
    Interleukin 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1698 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human il 2
    VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) TNF-α, ( b ) <t>IL-6,</t> ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p
    Recombinant Human Il 2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 1387 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human il 4
    NK-4 down-regulates secretion of TARC by NHDF stimulated with both <t>IL-4</t> and TNF-α. NHDF were stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α in the presence or absence (control) of varying concentrations of NK-4, dexamethasone or FK-506 (A) for 48 h at 37°C. In experiments comparing NK-4 with suplatast tosilate (B), NHDF were exposed to test articles for 15 min before stimulation with IL-4 and TNF-α. TARC levels in the cultures of NHDF without stimulation were below detectable limits. Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. ** p
    Recombinant Human Il 4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 986 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems pdgf bb
    Incubation of <t>TGF-β1</t> NAB in VSMCs increased the expression of lamin A in cocultured ECs, but <t>PDGF-BB</t> had no significant effect ( A ). NABs of both PDGF-BB and TGF-β1 incubation in VSMCs decreased expressions of LOX and phospho-ERK1/2 ( A
    Pdgf Bb, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 2186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems rmil 4
    Incubation of <t>TGF-β1</t> NAB in VSMCs increased the expression of lamin A in cocultured ECs, but <t>PDGF-BB</t> had no significant effect ( A ). NABs of both PDGF-BB and TGF-β1 incubation in VSMCs decreased expressions of LOX and phospho-ERK1/2 ( A
    Rmil 4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 420 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad recombinant proteins
    Incubation of <t>TGF-β1</t> NAB in VSMCs increased the expression of lamin A in cocultured ECs, but <t>PDGF-BB</t> had no significant effect ( A ). NABs of both PDGF-BB and TGF-β1 incubation in VSMCs decreased expressions of LOX and phospho-ERK1/2 ( A
    Recombinant Proteins, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 339 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Rhodomyrtone inhibits TNF-α+IL-17A-induced inflammatory responses in skin organ cultures. . Bars indicate mean ± SD, n = 5. Statistical significance determined by one-way ANOVA and Dunn’s multiple comparison test, indicated as* p

    Journal: PLoS ONE

    Article Title: The small molecule rhodomyrtone suppresses TNF-α and IL-17A-induced keratinocyte inflammatory responses: A potential new therapeutic for psoriasis

    doi: 10.1371/journal.pone.0205340

    Figure Lengend Snippet: Rhodomyrtone inhibits TNF-α+IL-17A-induced inflammatory responses in skin organ cultures. . Bars indicate mean ± SD, n = 5. Statistical significance determined by one-way ANOVA and Dunn’s multiple comparison test, indicated as* p

    Article Snippet: NHK were pretreated with rhodomyrtone (50–600 ng/ml) for 1 h before stimulation with recombinant human 20ng/ml IL-17A and 10 ng/ml TNF-α (both R & D Systems) for 24 h. Cells were harvested for RNA isolation in complete RLT buffer (Qiagen) or protein isolation in complete RIPA buffer.

    Techniques:

    Rhodomyrtone preferentially down-regulates transcripts involved in epidermal responses, myeloid leukocyte chemotaxis and inflammatory responses. Global transcriptome sequencing (RNA-seq) was performed on NHK treated with IL-17A+TNF for 24 h with or without 1 h pre-treatment with 400 ng/ml rhodomyrtone. Rhodomyrtone inhibited the expression of 579 of 1066 IL-17A/TNF-induced genes by NHK (2-fold change, TREAT p ] showed a decrease in the expression of IL-17/TNF induced transcripts area under curve = 0.46 (f) vs. 0.30 (h) and a decrease in genes overlapping with a plaque psoriasis gene signature, area under curve = 0.21 (g) vs. 0.07 (i).

    Journal: PLoS ONE

    Article Title: The small molecule rhodomyrtone suppresses TNF-α and IL-17A-induced keratinocyte inflammatory responses: A potential new therapeutic for psoriasis

    doi: 10.1371/journal.pone.0205340

    Figure Lengend Snippet: Rhodomyrtone preferentially down-regulates transcripts involved in epidermal responses, myeloid leukocyte chemotaxis and inflammatory responses. Global transcriptome sequencing (RNA-seq) was performed on NHK treated with IL-17A+TNF for 24 h with or without 1 h pre-treatment with 400 ng/ml rhodomyrtone. Rhodomyrtone inhibited the expression of 579 of 1066 IL-17A/TNF-induced genes by NHK (2-fold change, TREAT p ] showed a decrease in the expression of IL-17/TNF induced transcripts area under curve = 0.46 (f) vs. 0.30 (h) and a decrease in genes overlapping with a plaque psoriasis gene signature, area under curve = 0.21 (g) vs. 0.07 (i).

    Article Snippet: NHK were pretreated with rhodomyrtone (50–600 ng/ml) for 1 h before stimulation with recombinant human 20ng/ml IL-17A and 10 ng/ml TNF-α (both R & D Systems) for 24 h. Cells were harvested for RNA isolation in complete RLT buffer (Qiagen) or protein isolation in complete RIPA buffer.

    Techniques: Chemotaxis Assay, Sequencing, RNA Sequencing Assay, Expressing

    Rhodomyrtone decreases expression of inflammatory transcripts by TNF-α+IL-17A stimulated in skin organ cultures. Skin biopsies were pre-treated with 400 ng/ml rhodomyrtone 6 h before stimulation with 10 ng/ml TNF-α and 20 ng/ml IL-17A. Rhodomyrtone treatment decreases production of DEFB4 , IL1B , IL17C , IL36G , LCN2 , PI3 , S100A7 , and S100A8 transcripts as measured by qRT-PCR at 72 h. Statistical significance determined by one-way ANOVA and Dunn’s multiple comparison test, indicated as * p

    Journal: PLoS ONE

    Article Title: The small molecule rhodomyrtone suppresses TNF-α and IL-17A-induced keratinocyte inflammatory responses: A potential new therapeutic for psoriasis

    doi: 10.1371/journal.pone.0205340

    Figure Lengend Snippet: Rhodomyrtone decreases expression of inflammatory transcripts by TNF-α+IL-17A stimulated in skin organ cultures. Skin biopsies were pre-treated with 400 ng/ml rhodomyrtone 6 h before stimulation with 10 ng/ml TNF-α and 20 ng/ml IL-17A. Rhodomyrtone treatment decreases production of DEFB4 , IL1B , IL17C , IL36G , LCN2 , PI3 , S100A7 , and S100A8 transcripts as measured by qRT-PCR at 72 h. Statistical significance determined by one-way ANOVA and Dunn’s multiple comparison test, indicated as * p

    Article Snippet: NHK were pretreated with rhodomyrtone (50–600 ng/ml) for 1 h before stimulation with recombinant human 20ng/ml IL-17A and 10 ng/ml TNF-α (both R & D Systems) for 24 h. Cells were harvested for RNA isolation in complete RLT buffer (Qiagen) or protein isolation in complete RIPA buffer.

    Techniques: Expressing, Quantitative RT-PCR

    Rhodomyrtone dose dependently decreases production of CXCL1 , IL-36G , IL-36RN , MMP-13 , S100A7 , S100A8 , and S100A12 transcripts by IL-17A/TNF-α-stimulated primary human keratinocytes. Keratinocytes were pretreated with rhodomyrtone (50, 100, 200, 400, and 600 ng/ml) before stimulation with 20ng/ml IL-17A+10ng/ml TNF-α. The mRNA expression levels were measured at 24 h by qRT-PCR. Values are mean±SD of results from three independent experiments in triplicate. Statistical significance determined using one-way ANOVA and Dunnett’s multiple comparison test and noted **** p

    Journal: PLoS ONE

    Article Title: The small molecule rhodomyrtone suppresses TNF-α and IL-17A-induced keratinocyte inflammatory responses: A potential new therapeutic for psoriasis

    doi: 10.1371/journal.pone.0205340

    Figure Lengend Snippet: Rhodomyrtone dose dependently decreases production of CXCL1 , IL-36G , IL-36RN , MMP-13 , S100A7 , S100A8 , and S100A12 transcripts by IL-17A/TNF-α-stimulated primary human keratinocytes. Keratinocytes were pretreated with rhodomyrtone (50, 100, 200, 400, and 600 ng/ml) before stimulation with 20ng/ml IL-17A+10ng/ml TNF-α. The mRNA expression levels were measured at 24 h by qRT-PCR. Values are mean±SD of results from three independent experiments in triplicate. Statistical significance determined using one-way ANOVA and Dunnett’s multiple comparison test and noted **** p

    Article Snippet: NHK were pretreated with rhodomyrtone (50–600 ng/ml) for 1 h before stimulation with recombinant human 20ng/ml IL-17A and 10 ng/ml TNF-α (both R & D Systems) for 24 h. Cells were harvested for RNA isolation in complete RLT buffer (Qiagen) or protein isolation in complete RIPA buffer.

    Techniques: Expressing, Quantitative RT-PCR

    TNF-α and IL-17A drive psoriasis transcript expression in ex vivo skin organ cultures. To validate the use of cytokine-stimulated normal human skin biopsies as a model of skin inflammation, we treated biopsies with a combination of 10 ng/ml TNF-α and 20 ng/ml IL-17A for 12, 24, 48, and 72 h and monitored expression of DEFB4 , PI3 , LCN2 , S100A7 , and IL36G transcripts by qRT-PCR. Values are mean ± SD of results in triplicate from 3 donors. Statistical significance indicated by * p

    Journal: PLoS ONE

    Article Title: The small molecule rhodomyrtone suppresses TNF-α and IL-17A-induced keratinocyte inflammatory responses: A potential new therapeutic for psoriasis

    doi: 10.1371/journal.pone.0205340

    Figure Lengend Snippet: TNF-α and IL-17A drive psoriasis transcript expression in ex vivo skin organ cultures. To validate the use of cytokine-stimulated normal human skin biopsies as a model of skin inflammation, we treated biopsies with a combination of 10 ng/ml TNF-α and 20 ng/ml IL-17A for 12, 24, 48, and 72 h and monitored expression of DEFB4 , PI3 , LCN2 , S100A7 , and IL36G transcripts by qRT-PCR. Values are mean ± SD of results in triplicate from 3 donors. Statistical significance indicated by * p

    Article Snippet: NHK were pretreated with rhodomyrtone (50–600 ng/ml) for 1 h before stimulation with recombinant human 20ng/ml IL-17A and 10 ng/ml TNF-α (both R & D Systems) for 24 h. Cells were harvested for RNA isolation in complete RLT buffer (Qiagen) or protein isolation in complete RIPA buffer.

    Techniques: Expressing, Ex Vivo, Quantitative RT-PCR

    Rhodomyrtone inhibits IL-17A/TNF-α-induced inflammatory responses in keratinocytes. Primary human keratinocytes were pretreated with rhodomyrtone (50, 100, 200, 400, and 600 ng/ml) for 1 h before stimulation with 20 ng/ml IL-17A+10 ng/ml TNF-α. HBD-2 (a), and CXCL-1 (b), were assayed in conditioned media at 24 h by ELISA and found to be reduced in a dose-dependent manner in the presence of rhodomyrtone. Values are mean ± SD of results from three independent experiments in triplicate. Statistical significance determined using one-way ANOVA and Dunnett’s multiple comparison test and noted **** p

    Journal: PLoS ONE

    Article Title: The small molecule rhodomyrtone suppresses TNF-α and IL-17A-induced keratinocyte inflammatory responses: A potential new therapeutic for psoriasis

    doi: 10.1371/journal.pone.0205340

    Figure Lengend Snippet: Rhodomyrtone inhibits IL-17A/TNF-α-induced inflammatory responses in keratinocytes. Primary human keratinocytes were pretreated with rhodomyrtone (50, 100, 200, 400, and 600 ng/ml) for 1 h before stimulation with 20 ng/ml IL-17A+10 ng/ml TNF-α. HBD-2 (a), and CXCL-1 (b), were assayed in conditioned media at 24 h by ELISA and found to be reduced in a dose-dependent manner in the presence of rhodomyrtone. Values are mean ± SD of results from three independent experiments in triplicate. Statistical significance determined using one-way ANOVA and Dunnett’s multiple comparison test and noted **** p

    Article Snippet: NHK were pretreated with rhodomyrtone (50–600 ng/ml) for 1 h before stimulation with recombinant human 20ng/ml IL-17A and 10 ng/ml TNF-α (both R & D Systems) for 24 h. Cells were harvested for RNA isolation in complete RLT buffer (Qiagen) or protein isolation in complete RIPA buffer.

    Techniques: Enzyme-linked Immunosorbent Assay

    VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) TNF-α, ( b ) IL-6, ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p

    Journal: Vaccines

    Article Title: Virus-Like Particles Are a Superior Platform for Presenting M2e Epitopes to Prime Humoral and Cellular Immunity against Influenza Virus

    doi: 10.3390/vaccines6040066

    Figure Lengend Snippet: VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) TNF-α, ( b ) IL-6, ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p

    Article Snippet: Briefly, interleukin-6 (IL-6), IL-10, tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ) kits (eBioscience, San Diego, CA, USA) were used to determine the level of cytokines in the BALF and culture supernatants according to the manufacturer’s procedures.

    Techniques: In Vitro, Enzyme-linked Immunosorbent Assay

    Netrin-1 promotes inflammation resolution in diabetic corneal wound healing. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). Mouse cornea was collected and immunofluorescent stained with neutrophil marker Ly6G at 48 h and M2 macrophage markers CD68 (green fluorescence) and CD206 (red fluorescence) at 72 h after epithelial scrape ( A , 3 mice per group). Flow cytometry was performed to quantify the number and percentage of neutrophils (CD45 + CD11b + Ly6G+) ( B , 12 mice per group) at 48-hour post-wounded, M1 macrophages (CD45 + F4/80 + CD86+) and M2 macrophages (CD45 + F4/80 + CD206+) at 72-hour post-wounded. ( C , 12 mice per group at each detection) mRNA expression levels of iNOS and IL-12 (48 h after epithelial scrape), arginase-1 and IL-10 (72 h after epithelial scrape) were analyzed by RT-qPCR from the control and diabetic mouse corneas ( D , 6 mice per group); *p

    Journal: Scientific Reports

    Article Title: Netrin-1 promotes diabetic corneal wound healing through molecular mechanisms mediated via the adenosine 2B receptor

    doi: 10.1038/s41598-018-24506-9

    Figure Lengend Snippet: Netrin-1 promotes inflammation resolution in diabetic corneal wound healing. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). Mouse cornea was collected and immunofluorescent stained with neutrophil marker Ly6G at 48 h and M2 macrophage markers CD68 (green fluorescence) and CD206 (red fluorescence) at 72 h after epithelial scrape ( A , 3 mice per group). Flow cytometry was performed to quantify the number and percentage of neutrophils (CD45 + CD11b + Ly6G+) ( B , 12 mice per group) at 48-hour post-wounded, M1 macrophages (CD45 + F4/80 + CD86+) and M2 macrophages (CD45 + F4/80 + CD206+) at 72-hour post-wounded. ( C , 12 mice per group at each detection) mRNA expression levels of iNOS and IL-12 (48 h after epithelial scrape), arginase-1 and IL-10 (72 h after epithelial scrape) were analyzed by RT-qPCR from the control and diabetic mouse corneas ( D , 6 mice per group); *p

    Article Snippet: After the corneal epithelium scraped, diabetic mice were injected subconjunctivally with 50 ng netrin-1 (5 μ L/eye, R & D Systems, Minneapolis, MN) or equal phosphate buffer saline (PBS) as the vehicle control.

    Techniques: Mouse Assay, Injection, Staining, Marker, Fluorescence, Flow Cytometry, Cytometry, Expressing, Quantitative RT-PCR

    Netrin-1 promotes corneal epithelial wound healing in diabetic mice. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). The corneal epithelial wound defect was stained with fluorescein sodium at 24, 48, and 72 h after epithelial scrape ( A , 8 mice per group). The histogram of residual epithelial defect is presented as the percentage of the original wound area ( B ). *p

    Journal: Scientific Reports

    Article Title: Netrin-1 promotes diabetic corneal wound healing through molecular mechanisms mediated via the adenosine 2B receptor

    doi: 10.1038/s41598-018-24506-9

    Figure Lengend Snippet: Netrin-1 promotes corneal epithelial wound healing in diabetic mice. Corneal epithelium was scraped in control and diabetic mice with or without subconjunctival injection of netrin-1 (50 ng per eye). The corneal epithelial wound defect was stained with fluorescein sodium at 24, 48, and 72 h after epithelial scrape ( A , 8 mice per group). The histogram of residual epithelial defect is presented as the percentage of the original wound area ( B ). *p

    Article Snippet: After the corneal epithelium scraped, diabetic mice were injected subconjunctivally with 50 ng netrin-1 (5 μ L/eye, R & D Systems, Minneapolis, MN) or equal phosphate buffer saline (PBS) as the vehicle control.

    Techniques: Mouse Assay, Injection, Staining

    Netrin-1 promotes the migration, proliferation, and ERK and EGFR phosphorylation of corneal epithelial cells impaired by hyperglycemia. Confluent mouse corneal epithelial cells were wounded after incubation with 5 mM glucose, 30 mM mannose or 30 mM glucose. Cell migration was observed with or without 50 ng/mL netrin-1 treatment for 24 h ( A ). Migration rate was analyzed and represented as the percentage of primary wounding area ( B , n = 3 per group). Cell proliferation was measured by cell counting after 3 days of netrin-1 treatment ( C , n = 3 per group). The phosphorylated levels of ERK and EGFR were analyzed by Western blotting after 3 days incubation until monolayer formation. We administrated netrin-1 (50ng/ml) and after 8 or 15 minutes we collected the cells and then detected the p-ERK or p-EGFR expression by western blotting. ( D , n = 4 per group). The expression of p-ERK/p-EGFR were also confirmed by immunofluorescence staining in the netrin-1 injected diabetic mouse corneal epithelium 48 h after injury ( E , 3 mice per group). *p

    Journal: Scientific Reports

    Article Title: Netrin-1 promotes diabetic corneal wound healing through molecular mechanisms mediated via the adenosine 2B receptor

    doi: 10.1038/s41598-018-24506-9

    Figure Lengend Snippet: Netrin-1 promotes the migration, proliferation, and ERK and EGFR phosphorylation of corneal epithelial cells impaired by hyperglycemia. Confluent mouse corneal epithelial cells were wounded after incubation with 5 mM glucose, 30 mM mannose or 30 mM glucose. Cell migration was observed with or without 50 ng/mL netrin-1 treatment for 24 h ( A ). Migration rate was analyzed and represented as the percentage of primary wounding area ( B , n = 3 per group). Cell proliferation was measured by cell counting after 3 days of netrin-1 treatment ( C , n = 3 per group). The phosphorylated levels of ERK and EGFR were analyzed by Western blotting after 3 days incubation until monolayer formation. We administrated netrin-1 (50ng/ml) and after 8 or 15 minutes we collected the cells and then detected the p-ERK or p-EGFR expression by western blotting. ( D , n = 4 per group). The expression of p-ERK/p-EGFR were also confirmed by immunofluorescence staining in the netrin-1 injected diabetic mouse corneal epithelium 48 h after injury ( E , 3 mice per group). *p

    Article Snippet: After the corneal epithelium scraped, diabetic mice were injected subconjunctivally with 50 ng netrin-1 (5 μ L/eye, R & D Systems, Minneapolis, MN) or equal phosphate buffer saline (PBS) as the vehicle control.

    Techniques: Migration, Incubation, Cell Counting, Western Blot, Expressing, Immunofluorescence, Staining, Injection, Mouse Assay

    The A2BAR receptor mediates the promotion of netrin-1 on diabetic corneal wound healing. A2BAR receptor-specific antagonist or anti-UNC5B blocking antibody was injected subconjunctivally at 24 h before and 0, 24, and 48 h after the scrape of corneal epithelium in netrin-1 treated diabetic mice. Corneal epithelium defect was stained with fluorescein sodium and presented as the percentage of the original wound at 72 h after epithelial scraped ( A , 3 mice per group). Activation of ERK and EGFR in corneal epithelium were analyzed by western blotting at 48 h after epithelial scraped ( B , 3 mice per group). Neutrophils infiltration to the cornea and the cell numbers were examined by immunofluorescence staining ( C , 3 mice per group) and flow cytometry ( D , 12 mice per group) at 48 h after epithelial scraped. Macrophages infiltration to the cornea and the cell numbers were examined by immunofluorescence staining (C, 3 mice per group) and flow cytometry ( D , 12 mice per group of each detection) at 72 h after epithelial scraped. A2R anta: A2BAR antagonist; anti-U5B: anti-UNC5B blocking antibody. *p

    Journal: Scientific Reports

    Article Title: Netrin-1 promotes diabetic corneal wound healing through molecular mechanisms mediated via the adenosine 2B receptor

    doi: 10.1038/s41598-018-24506-9

    Figure Lengend Snippet: The A2BAR receptor mediates the promotion of netrin-1 on diabetic corneal wound healing. A2BAR receptor-specific antagonist or anti-UNC5B blocking antibody was injected subconjunctivally at 24 h before and 0, 24, and 48 h after the scrape of corneal epithelium in netrin-1 treated diabetic mice. Corneal epithelium defect was stained with fluorescein sodium and presented as the percentage of the original wound at 72 h after epithelial scraped ( A , 3 mice per group). Activation of ERK and EGFR in corneal epithelium were analyzed by western blotting at 48 h after epithelial scraped ( B , 3 mice per group). Neutrophils infiltration to the cornea and the cell numbers were examined by immunofluorescence staining ( C , 3 mice per group) and flow cytometry ( D , 12 mice per group) at 48 h after epithelial scraped. Macrophages infiltration to the cornea and the cell numbers were examined by immunofluorescence staining (C, 3 mice per group) and flow cytometry ( D , 12 mice per group of each detection) at 72 h after epithelial scraped. A2R anta: A2BAR antagonist; anti-U5B: anti-UNC5B blocking antibody. *p

    Article Snippet: After the corneal epithelium scraped, diabetic mice were injected subconjunctivally with 50 ng netrin-1 (5 μ L/eye, R & D Systems, Minneapolis, MN) or equal phosphate buffer saline (PBS) as the vehicle control.

    Techniques: Blocking Assay, Injection, Mouse Assay, Staining, Activation Assay, Western Blot, Immunofluorescence, Flow Cytometry, Cytometry

    Hyperglycemia downregulates netrin-1 expression in corneal epithelium. Normal and diabetic mouse corneal epithelium was collected before (as unwound group) or 48 h after epithelial scrape (as wound group). Netrin-1 expression was examined by using immunofluorescence staining ( A , 3 mice per group), RT-qPCR ( B , 9 mice per group) and ELISA ( C , 9 mice per group) with the age-matched normal mice as control. *p

    Journal: Scientific Reports

    Article Title: Netrin-1 promotes diabetic corneal wound healing through molecular mechanisms mediated via the adenosine 2B receptor

    doi: 10.1038/s41598-018-24506-9

    Figure Lengend Snippet: Hyperglycemia downregulates netrin-1 expression in corneal epithelium. Normal and diabetic mouse corneal epithelium was collected before (as unwound group) or 48 h after epithelial scrape (as wound group). Netrin-1 expression was examined by using immunofluorescence staining ( A , 3 mice per group), RT-qPCR ( B , 9 mice per group) and ELISA ( C , 9 mice per group) with the age-matched normal mice as control. *p

    Article Snippet: After the corneal epithelium scraped, diabetic mice were injected subconjunctivally with 50 ng netrin-1 (5 μ L/eye, R & D Systems, Minneapolis, MN) or equal phosphate buffer saline (PBS) as the vehicle control.

    Techniques: Expressing, Immunofluorescence, Staining, Mouse Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    The level of TNF-α protein increased in TOLF. A) Fold change of TNF-α protein in the ossified ligamentum flavum of TOLF. Proteins were extracted from the ossified ligamentum flavum (OS-Lig) and normal ligamentum flavum (Lig). The samples were then labeled with iTRAQ following the manufacture’s protocol. *: A star indicates statistical significance compared to control group with p

    Journal: PLoS ONE

    Article Title: The involvement and possible mechanism of pro-inflammatory tumor necrosis factor alpha (TNF-α) in thoracic ossification of the ligamentum flavum

    doi: 10.1371/journal.pone.0178986

    Figure Lengend Snippet: The level of TNF-α protein increased in TOLF. A) Fold change of TNF-α protein in the ossified ligamentum flavum of TOLF. Proteins were extracted from the ossified ligamentum flavum (OS-Lig) and normal ligamentum flavum (Lig). The samples were then labeled with iTRAQ following the manufacture’s protocol. *: A star indicates statistical significance compared to control group with p

    Article Snippet: Primary ligamentum flavum cells derived from TOLF patients were cultured and treated as indicated with recombinant human TNF-α (100 ng/ml; R & D Systems) for 24 hr.

    Techniques: Labeling

    Proportions of cells in S phase increased after TNF-α stimulation. Primary cells of the ossified ligamentum flavum were cultured in the absence of TNF-α (A) or presence of TNF-α (B). Flow cytometry assay were used to examine the cell cycle. The proportions of cells in S phase of cell cycle increased up to 17.06% after TNF-α stimulation.

    Journal: PLoS ONE

    Article Title: The involvement and possible mechanism of pro-inflammatory tumor necrosis factor alpha (TNF-α) in thoracic ossification of the ligamentum flavum

    doi: 10.1371/journal.pone.0178986

    Figure Lengend Snippet: Proportions of cells in S phase increased after TNF-α stimulation. Primary cells of the ossified ligamentum flavum were cultured in the absence of TNF-α (A) or presence of TNF-α (B). Flow cytometry assay were used to examine the cell cycle. The proportions of cells in S phase of cell cycle increased up to 17.06% after TNF-α stimulation.

    Article Snippet: Primary ligamentum flavum cells derived from TOLF patients were cultured and treated as indicated with recombinant human TNF-α (100 ng/ml; R & D Systems) for 24 hr.

    Techniques: Cell Culture, Flow Cytometry, Cytometry

    TNF-α activated the expressions of cyclin D1 and c-Myc. Primary ligamentum flavum cells derived from TOLF patients were cultured and treated with TNF-α (100 ng/ml) for 24 hr. Total RNA was isolated and measured by real time RT-PCR. The RNA level from the control group was normalized to a value of 1. Values were presented as the mean ±S.D. A paired t -test was performed comparing control and TNF-α group. *: A star indicates statistical significance compared to control group with p

    Journal: PLoS ONE

    Article Title: The involvement and possible mechanism of pro-inflammatory tumor necrosis factor alpha (TNF-α) in thoracic ossification of the ligamentum flavum

    doi: 10.1371/journal.pone.0178986

    Figure Lengend Snippet: TNF-α activated the expressions of cyclin D1 and c-Myc. Primary ligamentum flavum cells derived from TOLF patients were cultured and treated with TNF-α (100 ng/ml) for 24 hr. Total RNA was isolated and measured by real time RT-PCR. The RNA level from the control group was normalized to a value of 1. Values were presented as the mean ±S.D. A paired t -test was performed comparing control and TNF-α group. *: A star indicates statistical significance compared to control group with p

    Article Snippet: Primary ligamentum flavum cells derived from TOLF patients were cultured and treated as indicated with recombinant human TNF-α (100 ng/ml; R & D Systems) for 24 hr.

    Techniques: Derivative Assay, Cell Culture, Isolation, Quantitative RT-PCR

    TNF-α induced the expression of osteoblast genes in primary cells from TOLF. Primary ligamentum flavum cells were derived from TOLF patients. Primary cells were treated with 100 ng/ml TNF-α or as indicated for 24 hr. Total RNA was isolated and measured by real time RT-PCR. The RNA level from the control group was normalized to a value of 1. Values were presented as the mean ±S.D. A paired t -test was performed comparing control and TNF-α group. *: A star indicates statistical significance compared to control group with p

    Journal: PLoS ONE

    Article Title: The involvement and possible mechanism of pro-inflammatory tumor necrosis factor alpha (TNF-α) in thoracic ossification of the ligamentum flavum

    doi: 10.1371/journal.pone.0178986

    Figure Lengend Snippet: TNF-α induced the expression of osteoblast genes in primary cells from TOLF. Primary ligamentum flavum cells were derived from TOLF patients. Primary cells were treated with 100 ng/ml TNF-α or as indicated for 24 hr. Total RNA was isolated and measured by real time RT-PCR. The RNA level from the control group was normalized to a value of 1. Values were presented as the mean ±S.D. A paired t -test was performed comparing control and TNF-α group. *: A star indicates statistical significance compared to control group with p

    Article Snippet: Primary ligamentum flavum cells derived from TOLF patients were cultured and treated as indicated with recombinant human TNF-α (100 ng/ml; R & D Systems) for 24 hr.

    Techniques: Expressing, Derivative Assay, Isolation, Quantitative RT-PCR

    3.2. Association of extracellular histones, HMGB1, sTM, and IL-6 with ALI

    Journal: Medicine

    Article Title: Damage-associated molecular patterns in intensive care unit patients with acute liver injuries

    doi: 10.1097/MD.0000000000012780

    Figure Lengend Snippet: 3.2. Association of extracellular histones, HMGB1, sTM, and IL-6 with ALI

    Article Snippet: Plasma concentrations of soluble thrombomodulin (sTM) and interleukin-6 (IL-6) were determined using commercially available ELISA kits against human thrombomodulin and human IL-6, respectively (Quantikine; R & D Systems, Minneapolis, MN).

    Techniques:

    3.2. Association of extracellular histones, HMGB1, sTM, and IL-6 with ALI

    Journal: Medicine

    Article Title: Damage-associated molecular patterns in intensive care unit patients with acute liver injuries

    doi: 10.1097/MD.0000000000012780

    Figure Lengend Snippet: 3.2. Association of extracellular histones, HMGB1, sTM, and IL-6 with ALI

    Article Snippet: Plasma concentrations of soluble thrombomodulin (sTM) and interleukin-6 (IL-6) were determined using commercially available ELISA kits against human thrombomodulin and human IL-6, respectively (Quantikine; R & D Systems, Minneapolis, MN).

    Techniques:

    VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) TNF-α, ( b ) IL-6, ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p

    Journal: Vaccines

    Article Title: Virus-Like Particles Are a Superior Platform for Presenting M2e Epitopes to Prime Humoral and Cellular Immunity against Influenza Virus

    doi: 10.3390/vaccines6040066

    Figure Lengend Snippet: VLP effectively stimulates BMDCs in vitro to secrete cytokines. BMDCs were stimulated with M2e5x, M1 VLP, or protein (0.3, 1, 5 μg/mL) for 24 h. ( a – c ) The levels of cytokines ( a ) TNF-α, ( b ) IL-6, ( c ) IFN-γ were determined in the culture supernatants by ELISA assay. Un: Medium only, P: proteins, V: VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p

    Article Snippet: Briefly, interleukin-6 (IL-6), IL-10, tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ) kits (eBioscience, San Diego, CA, USA) were used to determine the level of cytokines in the BALF and culture supernatants according to the manufacturer’s procedures.

    Techniques: In Vitro, Enzyme-linked Immunosorbent Assay

    VLP is superior to protein in priming acute innate immune responses in vivo at the injection site. Balb/c mice ( n = 3) were i.p. injected with 200 μL of PBS, 10 μg 5xM2e VLP, or 25 μg 5xM2e protein. The levels of cytokine and chemokine were determined in sera and peritoneal exudates collected at the indicated times. The levels of cytokines (TNF-α, IL-6, IFN-γ) and chemokines (MCP-1, RANTES) in serum ( a ) and Peritoneal exudates ( b ). ( c – j ) Different phenotypic innate immune cells were determined in the cells collected from peritoneal exudates at 24 h post injection using flow cytometry. PRO: 5xM2e proteins, VLP: 5xM2e VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p

    Journal: Vaccines

    Article Title: Virus-Like Particles Are a Superior Platform for Presenting M2e Epitopes to Prime Humoral and Cellular Immunity against Influenza Virus

    doi: 10.3390/vaccines6040066

    Figure Lengend Snippet: VLP is superior to protein in priming acute innate immune responses in vivo at the injection site. Balb/c mice ( n = 3) were i.p. injected with 200 μL of PBS, 10 μg 5xM2e VLP, or 25 μg 5xM2e protein. The levels of cytokine and chemokine were determined in sera and peritoneal exudates collected at the indicated times. The levels of cytokines (TNF-α, IL-6, IFN-γ) and chemokines (MCP-1, RANTES) in serum ( a ) and Peritoneal exudates ( b ). ( c – j ) Different phenotypic innate immune cells were determined in the cells collected from peritoneal exudates at 24 h post injection using flow cytometry. PRO: 5xM2e proteins, VLP: 5xM2e VLP. The statistical significance was confirmed by one-way ANOVA and Dunnett’s multiple comparison test. Error bars indicate the means ± SEM of concentrations from individual animals. *; p

    Article Snippet: Briefly, interleukin-6 (IL-6), IL-10, tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ) kits (eBioscience, San Diego, CA, USA) were used to determine the level of cytokines in the BALF and culture supernatants according to the manufacturer’s procedures.

    Techniques: In Vivo, Injection, Mouse Assay, Flow Cytometry, Cytometry

    NK-4 down-regulates secretion of TARC by NHDF stimulated with both IL-4 and TNF-α. NHDF were stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α in the presence or absence (control) of varying concentrations of NK-4, dexamethasone or FK-506 (A) for 48 h at 37°C. In experiments comparing NK-4 with suplatast tosilate (B), NHDF were exposed to test articles for 15 min before stimulation with IL-4 and TNF-α. TARC levels in the cultures of NHDF without stimulation were below detectable limits. Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. ** p

    Journal: PLoS ONE

    Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

    doi: 10.1371/journal.pone.0199666

    Figure Lengend Snippet: NK-4 down-regulates secretion of TARC by NHDF stimulated with both IL-4 and TNF-α. NHDF were stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α in the presence or absence (control) of varying concentrations of NK-4, dexamethasone or FK-506 (A) for 48 h at 37°C. In experiments comparing NK-4 with suplatast tosilate (B), NHDF were exposed to test articles for 15 min before stimulation with IL-4 and TNF-α. TARC levels in the cultures of NHDF without stimulation were below detectable limits. Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. ** p

    Article Snippet: Recombinant human IL-4 was purchased from R & D Systems (Minneapolis, MN).

    Techniques:

    NK-4 selectively down-regulates IL-4 production by primary splenic T cells in response to anti-CD3ε mAb. BALB/c moue spleen cells (1.2 x 10 6 cells/well) were stimulated with 5 μg/ml anti-CD3ε mAb in the presence or absence (control) of varying concentrations of NK-4 for 48 h at 37°C in 96-well plates. Concentrations of IFN-γ (A) and IL-4 (B) in culture supernatants were measured by ELISA. The ratios of IFN-γ to IL-4 are shown (C). Results are the means ± S.D. of triplicate cultures. Results are representative of two independent experiments with similar results. ** p

    Journal: PLoS ONE

    Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

    doi: 10.1371/journal.pone.0199666

    Figure Lengend Snippet: NK-4 selectively down-regulates IL-4 production by primary splenic T cells in response to anti-CD3ε mAb. BALB/c moue spleen cells (1.2 x 10 6 cells/well) were stimulated with 5 μg/ml anti-CD3ε mAb in the presence or absence (control) of varying concentrations of NK-4 for 48 h at 37°C in 96-well plates. Concentrations of IFN-γ (A) and IL-4 (B) in culture supernatants were measured by ELISA. The ratios of IFN-γ to IL-4 are shown (C). Results are the means ± S.D. of triplicate cultures. Results are representative of two independent experiments with similar results. ** p

    Article Snippet: Recombinant human IL-4 was purchased from R & D Systems (Minneapolis, MN).

    Techniques: Enzyme-linked Immunosorbent Assay

    NK-4 inhibits the IL-4-driven alteration of cytokine expression profile in THP-1 cells. THP-1 cells were cultured with 4 ng/ml and 10 ng/ml IL-4 for 3 days at 37°C in RPMI1640 medium containing 1% FCS in 12-well culture plate (A to C). In a separate experiment, THP-1 cells were cultured with 10 ng/ml IL-4 in the presence or absence of varying concentrations of NK-4 (D to F). After pre-treatment with IL-4, THP-1 cells (1.2 x 10 5 cells/well) in fresh RPMI1640 medium containing 10% FCS were stimulated overnight with LPS and the levels of TNF-α and IL-10 in the culture supernatants were determined by specific ELISA. The ratios of TNF-α to IL-10 are shown (C and F). Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. * p

    Journal: PLoS ONE

    Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

    doi: 10.1371/journal.pone.0199666

    Figure Lengend Snippet: NK-4 inhibits the IL-4-driven alteration of cytokine expression profile in THP-1 cells. THP-1 cells were cultured with 4 ng/ml and 10 ng/ml IL-4 for 3 days at 37°C in RPMI1640 medium containing 1% FCS in 12-well culture plate (A to C). In a separate experiment, THP-1 cells were cultured with 10 ng/ml IL-4 in the presence or absence of varying concentrations of NK-4 (D to F). After pre-treatment with IL-4, THP-1 cells (1.2 x 10 5 cells/well) in fresh RPMI1640 medium containing 10% FCS were stimulated overnight with LPS and the levels of TNF-α and IL-10 in the culture supernatants were determined by specific ELISA. The ratios of TNF-α to IL-10 are shown (C and F). Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. * p

    Article Snippet: Recombinant human IL-4 was purchased from R & D Systems (Minneapolis, MN).

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

    NK-4 selectively down-regulates Th2 cytokine production by antigen-stimulated established Th2 cells. Th1 clone #4 (2.5 x 10 4 cells/well) were stimulated with OVA (200 μg/ml) and MMC-treated BALB/c mouse spleen cells (1.2 x 10 6 cells/well) in the presence or absence (control) of varying concentrations of NK-4 for 48 h at 37°C in 96-well plates (A). D10.G4.1 cells (2.5 x 10 4 cells/well) were stimulated with MMC-treated C57BL/6 mouse spleen cells (1.2 x 10 6 cells/well) (B and C). Concentrations of IFN-γ (A), IL-4 (B) and IL-5 (C) in culture supernatants were measured by ELISA. Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. ** p

    Journal: PLoS ONE

    Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

    doi: 10.1371/journal.pone.0199666

    Figure Lengend Snippet: NK-4 selectively down-regulates Th2 cytokine production by antigen-stimulated established Th2 cells. Th1 clone #4 (2.5 x 10 4 cells/well) were stimulated with OVA (200 μg/ml) and MMC-treated BALB/c mouse spleen cells (1.2 x 10 6 cells/well) in the presence or absence (control) of varying concentrations of NK-4 for 48 h at 37°C in 96-well plates (A). D10.G4.1 cells (2.5 x 10 4 cells/well) were stimulated with MMC-treated C57BL/6 mouse spleen cells (1.2 x 10 6 cells/well) (B and C). Concentrations of IFN-γ (A), IL-4 (B) and IL-5 (C) in culture supernatants were measured by ELISA. Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. ** p

    Article Snippet: Recombinant human IL-4 was purchased from R & D Systems (Minneapolis, MN).

    Techniques: Enzyme-linked Immunosorbent Assay

    NK-4 suppresses the STAT6 signaling pathway in NHDF stimulated with IL-4 and TNF-α. NHDF were grown in confluent monolayer cultures in 12-well plates and were stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α in the presence or absence of varying concentrations of NK-4 for 15 min. Phosphorylation of STAT6 was determined by immunoblotting whole-cell lysates using specific antibodies against the phosphorylated or total STAT6 protein. A representative blot is shown (A). The optical density ratio of phospho-STAT6 to total STAT6 is shown (B). Data from three independent experiments were combined and expressed as the means ± SD. ** p ).

    Journal: PLoS ONE

    Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

    doi: 10.1371/journal.pone.0199666

    Figure Lengend Snippet: NK-4 suppresses the STAT6 signaling pathway in NHDF stimulated with IL-4 and TNF-α. NHDF were grown in confluent monolayer cultures in 12-well plates and were stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α in the presence or absence of varying concentrations of NK-4 for 15 min. Phosphorylation of STAT6 was determined by immunoblotting whole-cell lysates using specific antibodies against the phosphorylated or total STAT6 protein. A representative blot is shown (A). The optical density ratio of phospho-STAT6 to total STAT6 is shown (B). Data from three independent experiments were combined and expressed as the means ± SD. ** p ).

    Article Snippet: Recombinant human IL-4 was purchased from R & D Systems (Minneapolis, MN).

    Techniques:

    NK-4 down-regulates secretion of eotaxin by NHDF in response to IL-4 and/or TNF-α. NHDF were stimulated with varying concentrations of IL-4 (A) or TNF-α (B) in the presence or absence (control) of 10 μM NK-4 for 48 h at 37°C. The effect of NK-4 on the secretion of eotaxin from NHDF stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α was assessed (C and D). Concentrations of eotaxin in culture supernatants were measured by ELISA (A to C). Cell numbers of NHDF were determined by cell counting kit-8 (D). Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. * p

    Journal: PLoS ONE

    Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

    doi: 10.1371/journal.pone.0199666

    Figure Lengend Snippet: NK-4 down-regulates secretion of eotaxin by NHDF in response to IL-4 and/or TNF-α. NHDF were stimulated with varying concentrations of IL-4 (A) or TNF-α (B) in the presence or absence (control) of 10 μM NK-4 for 48 h at 37°C. The effect of NK-4 on the secretion of eotaxin from NHDF stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α was assessed (C and D). Concentrations of eotaxin in culture supernatants were measured by ELISA (A to C). Cell numbers of NHDF were determined by cell counting kit-8 (D). Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. * p

    Article Snippet: Recombinant human IL-4 was purchased from R & D Systems (Minneapolis, MN).

    Techniques: Enzyme-linked Immunosorbent Assay, Cell Counting

    NK-4 selectively down-regulates Th2 cytokine production by anti-CD3ε mAb-stimulated established Th2 cells. Th1 clone #4 (A and B) and Th2 clone D10.G4.1 cells (C to E) (2.5 x 10 4 cells/well) were stimulated with immobilized anti-CD3ε mAb (8 μg/ml) in the presence or absence (control) of varying concentrations of NK-4 for 48 h at 37°C in 96-well plates. Concentrations of IFN-γ (A), IL-4 (C) and IL-5 (D) in culture supernatants were measured by ELISA. Cell numbers of #4 (B) and D10.G4.1 (E) were determined by cell counting kit-8. Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. * p

    Journal: PLoS ONE

    Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

    doi: 10.1371/journal.pone.0199666

    Figure Lengend Snippet: NK-4 selectively down-regulates Th2 cytokine production by anti-CD3ε mAb-stimulated established Th2 cells. Th1 clone #4 (A and B) and Th2 clone D10.G4.1 cells (C to E) (2.5 x 10 4 cells/well) were stimulated with immobilized anti-CD3ε mAb (8 μg/ml) in the presence or absence (control) of varying concentrations of NK-4 for 48 h at 37°C in 96-well plates. Concentrations of IFN-γ (A), IL-4 (C) and IL-5 (D) in culture supernatants were measured by ELISA. Cell numbers of #4 (B) and D10.G4.1 (E) were determined by cell counting kit-8. Results are the means ± S.D. of triplicate cultures. Results are representative of three independent experiments with similar results. * p

    Article Snippet: Recombinant human IL-4 was purchased from R & D Systems (Minneapolis, MN).

    Techniques: Enzyme-linked Immunosorbent Assay, Cell Counting

    Incubation of TGF-β1 NAB in VSMCs increased the expression of lamin A in cocultured ECs, but PDGF-BB had no significant effect ( A ). NABs of both PDGF-BB and TGF-β1 incubation in VSMCs decreased expressions of LOX and phospho-ERK1/2 ( A

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

    Article Title: PDGF-BB and TGF-?1 on cross-talk between endothelial and smooth muscle cells in vascular remodeling induced by low shear stress

    doi: 10.1073/pnas.1019219108

    Figure Lengend Snippet: Incubation of TGF-β1 NAB in VSMCs increased the expression of lamin A in cocultured ECs, but PDGF-BB had no significant effect ( A ). NABs of both PDGF-BB and TGF-β1 incubation in VSMCs decreased expressions of LOX and phospho-ERK1/2 ( A

    Article Snippet: Recombinant proteins of PDGF-BB and TGF-β1, NABs of PDGF-BB and TGF-β1, and Quantikine ELISA Kit for PDGF-BB and TGF-β1 were purchased from R & D Systems.

    Techniques: Incubation, Expressing

    PDGF-BB target siRNA transfection significantly decreased the secretions of PDGF-BB and TGF-β1 from ECs under LowSS conditions ( A ). PDGF-BB knockdown in ECs also markedly repressed the secretion of PDGF-BB and TGF-β1 from cocultured VSMCs

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

    Article Title: PDGF-BB and TGF-?1 on cross-talk between endothelial and smooth muscle cells in vascular remodeling induced by low shear stress

    doi: 10.1073/pnas.1019219108

    Figure Lengend Snippet: PDGF-BB target siRNA transfection significantly decreased the secretions of PDGF-BB and TGF-β1 from ECs under LowSS conditions ( A ). PDGF-BB knockdown in ECs also markedly repressed the secretion of PDGF-BB and TGF-β1 from cocultured VSMCs

    Article Snippet: Recombinant proteins of PDGF-BB and TGF-β1, NABs of PDGF-BB and TGF-β1, and Quantikine ELISA Kit for PDGF-BB and TGF-β1 were purchased from R & D Systems.

    Techniques: Transfection

    In ECs and VSMCs, LowSS induced the secretion of PDGF-BB and TGF-β1 at 12 and 24 h ( A and B ) and decreased the expression of lamin A and LOX at 6, 12, and 24 h ( C and D ). The phosphorylation of ERK1/2 was up-regulated by LowSS at 6 and 12 h in

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

    Article Title: PDGF-BB and TGF-?1 on cross-talk between endothelial and smooth muscle cells in vascular remodeling induced by low shear stress

    doi: 10.1073/pnas.1019219108

    Figure Lengend Snippet: In ECs and VSMCs, LowSS induced the secretion of PDGF-BB and TGF-β1 at 12 and 24 h ( A and B ) and decreased the expression of lamin A and LOX at 6, 12, and 24 h ( C and D ). The phosphorylation of ERK1/2 was up-regulated by LowSS at 6 and 12 h in

    Article Snippet: Recombinant proteins of PDGF-BB and TGF-β1, NABs of PDGF-BB and TGF-β1, and Quantikine ELISA Kit for PDGF-BB and TGF-β1 were purchased from R & D Systems.

    Techniques: Expressing