il 6 soluble receptor  (R&D Systems)

 
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    Name:
    Recombinant Human IL 6 Protein
    Description:
    The Recombinant Human IL 6 Protein from R D Systems is derived from E coli The Recombinant Human IL 6 Protein has been validated for the following applications Bioactivity
    Catalog Number:
    206-IL-010
    Price:
    229
    Category:
    Proteins and Enzymes
    Source:
    E. coli-derived Recombinant Human IL-6 Protein
    Applications:
    Bioactivity
    Purity:
    >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
    Conjugate:
    Unconjugated
    Size:
    10 ug
    Buy from Supplier


    Structured Review

    R&D Systems il 6 soluble receptor
    Recombinant Human IL 6 Protein
    The Recombinant Human IL 6 Protein from R D Systems is derived from E coli The Recombinant Human IL 6 Protein has been validated for the following applications Bioactivity
    https://www.bioz.com/result/il 6 soluble receptor/product/R&D Systems
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 6 soluble receptor - by Bioz Stars, 2021-06
    94/100 stars

    Images

    1) Product Images from "Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells"

    Article Title: Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

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

    doi: 10.1152/ajplung.00025.2019

    Effects of red blood cell (RBC) supernatants (SUP) on LPS or TNF-α-induced inflammatory response of human pulmonary artery endothelial cells (HPAECs). A and B : HPAECs were treated with LPS ( A ) or TNF-α ( B ) for 1 h followed by addition of fresh RBC supernatants (1:100 dilution) for 16 to 20 h and ELISA analysis of the markers of inflammation: soluble ICAM-1 (sICAM-1), IL-8, and IL-6; n = 3. * P
    Figure Legend Snippet: Effects of red blood cell (RBC) supernatants (SUP) on LPS or TNF-α-induced inflammatory response of human pulmonary artery endothelial cells (HPAECs). A and B : HPAECs were treated with LPS ( A ) or TNF-α ( B ) for 1 h followed by addition of fresh RBC supernatants (1:100 dilution) for 16 to 20 h and ELISA analysis of the markers of inflammation: soluble ICAM-1 (sICAM-1), IL-8, and IL-6; n = 3. * P

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Effects of red blood cell (RBC) supernatants (SUP) on inflammatory response by human pulmonary artery endothelial cells (HPAECs). HPAECs were challenged with RBC supernatants from fresh and old stored RBCs at 1:50 and 1:100 dilutions, or cells were stimulated with LPS as a positive control. A – C : after 16–20 h, the cultured medium was examined by ELISA for inflammatory markers: soluble ICAM-1 (sICAM-1) ( A ), IL-8 ( B ), and IL-6 ( C ); n = 5. * P
    Figure Legend Snippet: Effects of red blood cell (RBC) supernatants (SUP) on inflammatory response by human pulmonary artery endothelial cells (HPAECs). HPAECs were challenged with RBC supernatants from fresh and old stored RBCs at 1:50 and 1:100 dilutions, or cells were stimulated with LPS as a positive control. A – C : after 16–20 h, the cultured medium was examined by ELISA for inflammatory markers: soluble ICAM-1 (sICAM-1) ( A ), IL-8 ( B ), and IL-6 ( C ); n = 5. * P

    Techniques Used: Positive Control, Cell Culture, Enzyme-linked Immunosorbent Assay

    2) Product Images from "Interleukin-17 Enhancement of the Interleukin-6 Signaling Cascade in Astrocytes"

    Article Title: Interleukin-17 Enhancement of the Interleukin-6 Signaling Cascade in Astrocytes

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

    doi: 10.4049/jimmunol.1000142

    IL-17 Enhances IL-6-mediated SOCS3 Expression in Primary Astrocytes, and SOCS3 is a Negative Regulator of the Synergistic Effect of IL-6 and IL-17 A , Astrocytes were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 2 - 8 h, and levels of SOCS3 mRNA expression were determined by QRT-PCR. B and C, Astrocytes were transfected with SOCS3 siRNA (100 nM) or siRNA control (100 nM) for 48 h. Transfected cells were then treated with medium (UN) or IL-6/R plus IL-17 for 4 h. Levels of SOCS3 (B) and IL-6 (C) mRNA expression was determined by QRT-PCR. D, Astrocytes were transfected with SOCS3 siRNA or siRNA control for 48 h. Transfected cells were then treated with medium (UN) or IL-6/R plus IL-17 for 24 h, and supernatants analyzed for IL-6 protein by ELISA. E, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, cells were harvested. Digestion of genomic DNA distinguishes the full-length (fl) and excised alleles ( Δ ). F, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, the cells were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 4 h, and levels of IL-6 mRNA expression determined by QRT-PCR. G, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, cells were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 16 or 24 h, and supernatants analyzed for IL-6 protein by ELISA. All data are the mean ± SD of three experiments. *, p ≤ 0.05;**, p ≤ 0.01.
    Figure Legend Snippet: IL-17 Enhances IL-6-mediated SOCS3 Expression in Primary Astrocytes, and SOCS3 is a Negative Regulator of the Synergistic Effect of IL-6 and IL-17 A , Astrocytes were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 2 - 8 h, and levels of SOCS3 mRNA expression were determined by QRT-PCR. B and C, Astrocytes were transfected with SOCS3 siRNA (100 nM) or siRNA control (100 nM) for 48 h. Transfected cells were then treated with medium (UN) or IL-6/R plus IL-17 for 4 h. Levels of SOCS3 (B) and IL-6 (C) mRNA expression was determined by QRT-PCR. D, Astrocytes were transfected with SOCS3 siRNA or siRNA control for 48 h. Transfected cells were then treated with medium (UN) or IL-6/R plus IL-17 for 24 h, and supernatants analyzed for IL-6 protein by ELISA. E, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, cells were harvested. Digestion of genomic DNA distinguishes the full-length (fl) and excised alleles ( Δ ). F, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, the cells were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 4 h, and levels of IL-6 mRNA expression determined by QRT-PCR. G, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, cells were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 16 or 24 h, and supernatants analyzed for IL-6 protein by ELISA. All data are the mean ± SD of three experiments. *, p ≤ 0.05;**, p ≤ 0.01.

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay, Infection, Incubation

    Proposed Model of IL-17 Enhancement of the IL-6 Signaling Cascade in Astrocytes A, IL-6/R and IL-17 activate the NF-κB and MAPK pathway, which then induce IL-6 gene expression. The synergistic effect of these two mediators depends on NF-κB, p38 and JNK MAPK activity. Activated NF-κB p65, c-Fos and c-Jun bind to the IL-6 promoter. Concurrent with NF-κB and MAPK recruitment, IL-6/R and IL-17 leads to the recruitment of coactivators CBP and p300, modifications in AcH3 and AcH4, and recruitment of RNA Pol II to the IL-6 promoter, which results in transcriptional activation of the IL-6 gene. IL-17 can also enhance IL-6/R induced SOCS3 expression, and SOCS3 inhibits IL-6/R plus IL-17-induced NF-κB and MAPK activation, which results in a reduction of IL-6 gene expression in astrocytes. B , Naive CD4 + T cells, after activation by signaling through the T cell receptor and co-stimulatory molecules, can differentiate into Th17 cells in the presence of IL-6, TGF-β, IL-1 and IL-23. IL-17 together with IL-6/R triggers a positive-feedforward loop of IL-6 expression in astrocytes, which may also influence Th17 cell differentiation. SOCS3 participates in these processes as a negative feedback regulator. See text for details.
    Figure Legend Snippet: Proposed Model of IL-17 Enhancement of the IL-6 Signaling Cascade in Astrocytes A, IL-6/R and IL-17 activate the NF-κB and MAPK pathway, which then induce IL-6 gene expression. The synergistic effect of these two mediators depends on NF-κB, p38 and JNK MAPK activity. Activated NF-κB p65, c-Fos and c-Jun bind to the IL-6 promoter. Concurrent with NF-κB and MAPK recruitment, IL-6/R and IL-17 leads to the recruitment of coactivators CBP and p300, modifications in AcH3 and AcH4, and recruitment of RNA Pol II to the IL-6 promoter, which results in transcriptional activation of the IL-6 gene. IL-17 can also enhance IL-6/R induced SOCS3 expression, and SOCS3 inhibits IL-6/R plus IL-17-induced NF-κB and MAPK activation, which results in a reduction of IL-6 gene expression in astrocytes. B , Naive CD4 + T cells, after activation by signaling through the T cell receptor and co-stimulatory molecules, can differentiate into Th17 cells in the presence of IL-6, TGF-β, IL-1 and IL-23. IL-17 together with IL-6/R triggers a positive-feedforward loop of IL-6 expression in astrocytes, which may also influence Th17 cell differentiation. SOCS3 participates in these processes as a negative feedback regulator. See text for details.

    Techniques Used: Expressing, Activity Assay, Activation Assay, Cell Differentiation

    IL-17 Synergizes with IL-6/R for Activation of the NF-κB and MAPK Pathways A , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 15 and 30 min, and then cell lysates were immunoblotted with antibodies against IκBα and GAPDH. The basal level of the untreated sample was set at 100, and the percentage change of IκBα upon IL-6/R, IL-17 or IL-6/R + IL-17 treatment compared with the basal value. B , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 30 and 60 min, and then cell lysates were immunoblotted with antibodies against phospho-p65 Ser 536, p65 and GAPDH. C , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 15 and 30 min, and then cell lysates were immunoblotted with antibodies against phospho-ERK1/2, ERK1/2 and GAPDH. D , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R and IL-17 for 30 min and 60 min, and then cell lysates were immunoblotted with antibodies against phospho-p38, p38 and GAPDH. E , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 30 and 60 min, and then cell lysates were immunoblotted with antibodies against phospho-JNK, JNK and GAPDH. The basal level of the untreated sample was set at 1.0 and fold activation upon IL-6/R, IL-17 or IL-6/R + IL-17 treatment was compared with that value ( B , C , D and E ). Representative of at least three experiments.
    Figure Legend Snippet: IL-17 Synergizes with IL-6/R for Activation of the NF-κB and MAPK Pathways A , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 15 and 30 min, and then cell lysates were immunoblotted with antibodies against IκBα and GAPDH. The basal level of the untreated sample was set at 100, and the percentage change of IκBα upon IL-6/R, IL-17 or IL-6/R + IL-17 treatment compared with the basal value. B , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 30 and 60 min, and then cell lysates were immunoblotted with antibodies against phospho-p65 Ser 536, p65 and GAPDH. C , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 15 and 30 min, and then cell lysates were immunoblotted with antibodies against phospho-ERK1/2, ERK1/2 and GAPDH. D , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R and IL-17 for 30 min and 60 min, and then cell lysates were immunoblotted with antibodies against phospho-p38, p38 and GAPDH. E , Astrocytes were incubated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 30 and 60 min, and then cell lysates were immunoblotted with antibodies against phospho-JNK, JNK and GAPDH. The basal level of the untreated sample was set at 1.0 and fold activation upon IL-6/R, IL-17 or IL-6/R + IL-17 treatment was compared with that value ( B , C , D and E ). Representative of at least three experiments.

    Techniques Used: Activation Assay, Incubation

    IL-17 and IL-6/R Induction of IL-6 Depends on NF-κB, p38 and JNK MAPK Activity A , Astrocytes were cultured in absence or presence of IL-17 followed by IL-6/R treatment for 4 h. Actinomycin D (5 ng/ml) was then added, and cells harvested at 0, 30, 60, 120, and 240 min after addition. The abundance of IL-6 mRNA was determined by QRT-PCR. B , C , DMSO vehicle, BAY 11 (5 μM), U0126 (10 μM), SB203580 (10 μM) or JNKi II (10 μM) were added to cultures 1 h before cytokine addition, and then astrocytes were incubated with medium, IL-6/R, IL-17 or IL-6/R plus IL-17 for 4 h. Levels of IL-6 mRNA expression were determined by QRT-PCR. All data are the mean ± SD of three experiments. **, p ≤ 0.01; NS = not significant.
    Figure Legend Snippet: IL-17 and IL-6/R Induction of IL-6 Depends on NF-κB, p38 and JNK MAPK Activity A , Astrocytes were cultured in absence or presence of IL-17 followed by IL-6/R treatment for 4 h. Actinomycin D (5 ng/ml) was then added, and cells harvested at 0, 30, 60, 120, and 240 min after addition. The abundance of IL-6 mRNA was determined by QRT-PCR. B , C , DMSO vehicle, BAY 11 (5 μM), U0126 (10 μM), SB203580 (10 μM) or JNKi II (10 μM) were added to cultures 1 h before cytokine addition, and then astrocytes were incubated with medium, IL-6/R, IL-17 or IL-6/R plus IL-17 for 4 h. Levels of IL-6 mRNA expression were determined by QRT-PCR. All data are the mean ± SD of three experiments. **, p ≤ 0.01; NS = not significant.

    Techniques Used: Activity Assay, Cell Culture, Quantitative RT-PCR, Incubation, Expressing

    IL-6/R plus IL-17 Enhances Recruitment of p65, P-p65, c-Jun, c-Fos, CBP, p300, and RNA Pol II to the IL-6 Promoter A, Primary astrocytes were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 90 min, and then cells were cross-linked with formaldehyde. Soluble chromatin was subjected to immunoprecipitation with Abs against p65, P-p65, c-Fos, c-Jun, or normal rabbit IgG. PCR analysis of the positive control (input) indicates that soluble chromatin samples obtained from each time point had equal amounts of chromatin fragments containing the IL-6 promoter. B, Primary astrocytes were treated as above. Soluble chromatin was subjected to immunoprecipitation with Abs against histone acetylation (Ac-H3 and Ac-H4), p300, CBP, RNA Pol II or normal rabbit IgG. The basal level of the untreated sample was set at 1.0 and fold activation upon IL-6/R, IL-17 or IL-6/R plus IL-17 treatment was compared with that value. Representative of at least three experiments.
    Figure Legend Snippet: IL-6/R plus IL-17 Enhances Recruitment of p65, P-p65, c-Jun, c-Fos, CBP, p300, and RNA Pol II to the IL-6 Promoter A, Primary astrocytes were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 90 min, and then cells were cross-linked with formaldehyde. Soluble chromatin was subjected to immunoprecipitation with Abs against p65, P-p65, c-Fos, c-Jun, or normal rabbit IgG. PCR analysis of the positive control (input) indicates that soluble chromatin samples obtained from each time point had equal amounts of chromatin fragments containing the IL-6 promoter. B, Primary astrocytes were treated as above. Soluble chromatin was subjected to immunoprecipitation with Abs against histone acetylation (Ac-H3 and Ac-H4), p300, CBP, RNA Pol II or normal rabbit IgG. The basal level of the untreated sample was set at 1.0 and fold activation upon IL-6/R, IL-17 or IL-6/R plus IL-17 treatment was compared with that value. Representative of at least three experiments.

    Techniques Used: Immunoprecipitation, Polymerase Chain Reaction, Positive Control, Activation Assay

    Enhanced Activation of NF-κB and MAPK Pathways in SOCS3 Deficient Astrocytes To evaluate NF-κB and MAPK activation in the absence or presence of SOCS3, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, cells were treated with medium (UN) or IL-6/R plus IL-17 for 15, 30, 60 or 120 min, and then cell lysates were immunoblotted with antibodies against phospho-p65 Ser 536, p65, phospho-p38, p38, phospho-JNK, JNK or GAPDH. The basal level of the untreated GFP infected sample was set at 1.0 and fold activation upon IL-6/R plus IL-17 treatment compared with that value. Representative of at least three experiments.
    Figure Legend Snippet: Enhanced Activation of NF-κB and MAPK Pathways in SOCS3 Deficient Astrocytes To evaluate NF-κB and MAPK activation in the absence or presence of SOCS3, SOCS3 floxed astrocytes were infected with GFP as control or GFP-Cre for deletion. After 48 h in culture, cells were treated with medium (UN) or IL-6/R plus IL-17 for 15, 30, 60 or 120 min, and then cell lysates were immunoblotted with antibodies against phospho-p65 Ser 536, p65, phospho-p38, p38, phospho-JNK, JNK or GAPDH. The basal level of the untreated GFP infected sample was set at 1.0 and fold activation upon IL-6/R plus IL-17 treatment compared with that value. Representative of at least three experiments.

    Techniques Used: Activation Assay, Infection

    IL-17 Enhances IL-6/sIL-6R-mediated IL-6 Expression in Primary Astrocytes A , IL-17RA and IL-17RC mRNA expression was determined by RT-PCR in RAW264.7 cells (positive control), and primary astrocytes, in duplicate. B , Primary astrocytes were treated with medium (UN), IL-6/R (IL-6, 10 ng/ml and sIL-6R, 25 ng/ml), IL-17 (25 ng/ml) or IL-6/R plus IL-17 for up to 24 h, and levels of IL-6 and GAPDH mRNA expression were determined by RT-PCR and QRT-PCR. C , Primary astrocytes were treated with medium (UN), IL-6/R, different concentrations of IL-17 (1-50 ng/ml) or IL-6/R + IL-17 (1-50 ng/ml) for 4 h, and levels of IL-6 mRNA expression were determined by QRT-PCR. D , Primary astrocytes were treated with medium (UN), IL-6, sIL-6R or IL-17 in various combinations for 4 h, and levels of IL-6 mRNA expression were determined by QRT-PCR. E , Primary astrocytes were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 2 - 48 h, and supernatants were analyzed for IL-6 protein using ELISA. Experiments shown are representative of at least three experiments. All data are the mean ± SD of three experiments. *, p ≤0.05; **, p ≤0.01.
    Figure Legend Snippet: IL-17 Enhances IL-6/sIL-6R-mediated IL-6 Expression in Primary Astrocytes A , IL-17RA and IL-17RC mRNA expression was determined by RT-PCR in RAW264.7 cells (positive control), and primary astrocytes, in duplicate. B , Primary astrocytes were treated with medium (UN), IL-6/R (IL-6, 10 ng/ml and sIL-6R, 25 ng/ml), IL-17 (25 ng/ml) or IL-6/R plus IL-17 for up to 24 h, and levels of IL-6 and GAPDH mRNA expression were determined by RT-PCR and QRT-PCR. C , Primary astrocytes were treated with medium (UN), IL-6/R, different concentrations of IL-17 (1-50 ng/ml) or IL-6/R + IL-17 (1-50 ng/ml) for 4 h, and levels of IL-6 mRNA expression were determined by QRT-PCR. D , Primary astrocytes were treated with medium (UN), IL-6, sIL-6R or IL-17 in various combinations for 4 h, and levels of IL-6 mRNA expression were determined by QRT-PCR. E , Primary astrocytes were treated with medium (UN), IL-6/R, IL-17 or IL-6/R plus IL-17 for 2 - 48 h, and supernatants were analyzed for IL-6 protein using ELISA. Experiments shown are representative of at least three experiments. All data are the mean ± SD of three experiments. *, p ≤0.05; **, p ≤0.01.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    3) Product Images from "Astroglial expression of the P-glycoprotein is controlled by intracellular CNTF"

    Article Title: Astroglial expression of the P-glycoprotein is controlled by intracellular CNTF

    Journal: BMC Cell Biology

    doi: 10.1186/1471-2121-3-20

    Potentiation of IL-6 effects on P-gp expression by adding its specific α receptor subunits. Addition of soluble IL-6R induced a major potentiation of rmIL-6 whatever the concentration of the factor used.
    Figure Legend Snippet: Potentiation of IL-6 effects on P-gp expression by adding its specific α receptor subunits. Addition of soluble IL-6R induced a major potentiation of rmIL-6 whatever the concentration of the factor used.

    Techniques Used: Expressing, Concentration Assay

    Effect of cytokines of the IL-6 family on P-gp intracellular content in CNTF-/- astrocytes. Only rrCNTF (250 ng/ml) provoked a significant increase in P-gp intracellular content (+43.8 % ± 20, factorial ANOVA significant at 95%, t-test p
    Figure Legend Snippet: Effect of cytokines of the IL-6 family on P-gp intracellular content in CNTF-/- astrocytes. Only rrCNTF (250 ng/ml) provoked a significant increase in P-gp intracellular content (+43.8 % ± 20, factorial ANOVA significant at 95%, t-test p

    Techniques Used:

    Related Articles

    Recombinant:

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    Activity Assay:

    Article Title: Central Glucocorticoid Receptors Modulate the Expression and Function of Spinal NMDA Receptors after Peripheral Nerve Injury
    Article Snippet: The antibody was raised in goat (goat against rat), affinity purified, and diluted in PBS according to the manufacturer's information sheet. .. This antibody, showing < 1% cross-reactivity with recombinant human IL-6, was selected for its ability to neutralize the biological activity of recombinant rat IL-6, and the neutralization dose50 is 0.03-0.09 ng/μl (0.03-0.09 μg/ml) (R & D Systems). .. There was 0.5 μg of IL-6 IgG in 10 μl, and a goat IgG, diluted in the same solution for IL-6 IgG with the same concentration, was used as control serum.

    Neutralization:

    Article Title: Central Glucocorticoid Receptors Modulate the Expression and Function of Spinal NMDA Receptors after Peripheral Nerve Injury
    Article Snippet: The antibody was raised in goat (goat against rat), affinity purified, and diluted in PBS according to the manufacturer's information sheet. .. This antibody, showing < 1% cross-reactivity with recombinant human IL-6, was selected for its ability to neutralize the biological activity of recombinant rat IL-6, and the neutralization dose50 is 0.03-0.09 ng/μl (0.03-0.09 μg/ml) (R & D Systems). .. There was 0.5 μg of IL-6 IgG in 10 μl, and a goat IgG, diluted in the same solution for IL-6 IgG with the same concentration, was used as control serum.

    Cell Culture:

    Article Title: Discovery of a highly selective JAK3 inhibitor for the treatment of rheumatoid arthritis
    Article Snippet: Raw peripheral blood mononuclear cells (PBMCs; All Cells) were isolated from the buffy coats of healthy volunteers using density gradient centrifugation on Lymphoprep. .. Cells were cultured in complete RPMI 1640 medium (containing 10% foetal bovine serum, 100 mg/ml streptomycin and 100 U/ml penicillin) plus 10 μg/ml lectin phytohemagglutinin (PHA) for 3 days and then treated with either recombinant human IL-6 (400 ng/ml; R & D Systems), recombinant human IL-2 (100 ng/ml; R & D Systems), or recombinant human GM-CSF (50 ng/ml; Pepro-Tech) at 37 °C for 20 min. To terminate the stimulation, cells were fixed with Lyse/Fix Buffer and then incubated with 100% methanol for 30 minutes; cells were incubated with anti-pSTAT3 and anti-CD4 Abs, or anti-pSTAT5 and anti-CD4 Abs (all Abs were from BD Biosciences) at 4 °C overnight, washed twice with PBS, and analysed with an FACS Canto II flow cytometer. .. THP-1 cells (ATCC TIB-202) were preincubated with the compound at 37 °C for 1 h, incubated with IL-4 (10 ng/ml) at 37 °C for 60 min, and processed for Western blotting analysis.

    Incubation:

    Article Title: Discovery of a highly selective JAK3 inhibitor for the treatment of rheumatoid arthritis
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    FACS:

    Article Title: Discovery of a highly selective JAK3 inhibitor for the treatment of rheumatoid arthritis
    Article Snippet: Raw peripheral blood mononuclear cells (PBMCs; All Cells) were isolated from the buffy coats of healthy volunteers using density gradient centrifugation on Lymphoprep. .. Cells were cultured in complete RPMI 1640 medium (containing 10% foetal bovine serum, 100 mg/ml streptomycin and 100 U/ml penicillin) plus 10 μg/ml lectin phytohemagglutinin (PHA) for 3 days and then treated with either recombinant human IL-6 (400 ng/ml; R & D Systems), recombinant human IL-2 (100 ng/ml; R & D Systems), or recombinant human GM-CSF (50 ng/ml; Pepro-Tech) at 37 °C for 20 min. To terminate the stimulation, cells were fixed with Lyse/Fix Buffer and then incubated with 100% methanol for 30 minutes; cells were incubated with anti-pSTAT3 and anti-CD4 Abs, or anti-pSTAT5 and anti-CD4 Abs (all Abs were from BD Biosciences) at 4 °C overnight, washed twice with PBS, and analysed with an FACS Canto II flow cytometer. .. THP-1 cells (ATCC TIB-202) were preincubated with the compound at 37 °C for 1 h, incubated with IL-4 (10 ng/ml) at 37 °C for 60 min, and processed for Western blotting analysis.

    Flow Cytometry:

    Article Title: Discovery of a highly selective JAK3 inhibitor for the treatment of rheumatoid arthritis
    Article Snippet: Raw peripheral blood mononuclear cells (PBMCs; All Cells) were isolated from the buffy coats of healthy volunteers using density gradient centrifugation on Lymphoprep. .. Cells were cultured in complete RPMI 1640 medium (containing 10% foetal bovine serum, 100 mg/ml streptomycin and 100 U/ml penicillin) plus 10 μg/ml lectin phytohemagglutinin (PHA) for 3 days and then treated with either recombinant human IL-6 (400 ng/ml; R & D Systems), recombinant human IL-2 (100 ng/ml; R & D Systems), or recombinant human GM-CSF (50 ng/ml; Pepro-Tech) at 37 °C for 20 min. To terminate the stimulation, cells were fixed with Lyse/Fix Buffer and then incubated with 100% methanol for 30 minutes; cells were incubated with anti-pSTAT3 and anti-CD4 Abs, or anti-pSTAT5 and anti-CD4 Abs (all Abs were from BD Biosciences) at 4 °C overnight, washed twice with PBS, and analysed with an FACS Canto II flow cytometer. .. THP-1 cells (ATCC TIB-202) were preincubated with the compound at 37 °C for 1 h, incubated with IL-4 (10 ng/ml) at 37 °C for 60 min, and processed for Western blotting analysis.

    Cytometry:

    Article Title: Discovery of a highly selective JAK3 inhibitor for the treatment of rheumatoid arthritis
    Article Snippet: Raw peripheral blood mononuclear cells (PBMCs; All Cells) were isolated from the buffy coats of healthy volunteers using density gradient centrifugation on Lymphoprep. .. Cells were cultured in complete RPMI 1640 medium (containing 10% foetal bovine serum, 100 mg/ml streptomycin and 100 U/ml penicillin) plus 10 μg/ml lectin phytohemagglutinin (PHA) for 3 days and then treated with either recombinant human IL-6 (400 ng/ml; R & D Systems), recombinant human IL-2 (100 ng/ml; R & D Systems), or recombinant human GM-CSF (50 ng/ml; Pepro-Tech) at 37 °C for 20 min. To terminate the stimulation, cells were fixed with Lyse/Fix Buffer and then incubated with 100% methanol for 30 minutes; cells were incubated with anti-pSTAT3 and anti-CD4 Abs, or anti-pSTAT5 and anti-CD4 Abs (all Abs were from BD Biosciences) at 4 °C overnight, washed twice with PBS, and analysed with an FACS Canto II flow cytometer. .. THP-1 cells (ATCC TIB-202) were preincubated with the compound at 37 °C for 1 h, incubated with IL-4 (10 ng/ml) at 37 °C for 60 min, and processed for Western blotting analysis.

    Concentration Assay:

    Article Title: NFAT1-regulated IL6 signalling contributes to aggressive phenotypes of glioma
    Article Snippet: The multi-lineage differentiation capacity of GSCs was examined using anti-glial fibrillary acidic protein (GFAP, ab7260, Abcam) and anti-β III tubulin (ab78078, Abcam). .. Recombinant human IL-6 (R & D Systems, Minneapolis, MN, USA) was dissolved in sterile PBS (HyClone) at 100 μg/ml according to the manufacturer’s instructions, followed by different concentration preparations for glioma cell treatment. .. Real-time PCR Mini-BEST Universal RNA Extraction kit (TaKaRa, Kyoto, Japan) was used to isolate total RNA according to manufacturer’s instructions.

    Immunofluorescence:

    Article Title: IL-27 mediates HLA class I up-regulation, which can be inhibited by the IL-6 pathway, in HLA-deficient Small Cell Lung Cancer cells
    Article Snippet: .. For immunofluorescence and QRT-PCR analyses, cells were seeded in 24-well plates in culture medium at 5 × 104 cells/well and different cytokines were added: IFN-γ (1000 IU/ml, PeproTech, 300–02), IL-27 (100 ng/ml R & D System, 2526-IL-010), IL-6 (50 ng/ml R & D System 206-IL-010) or recombinant human IL-6Rα/IL-6 chimera [sIL-6R/IL-6] (50 ng/ml R & D System 8954-SR-025). .. Treatments were carried out for 48 h. For the analysis of tyrosine-phosphorylated STAT proteins, 1 × 105 SCLC cells were incubated in a test tube at 37 °C with or without 50 ng/ ml of IL-27, 20 ng/ml of IL-6, 40 ng/ml of sIL-6R/IL-6 in 0.5 ml of medium for the 10, 30 or 60 min time points.

    Quantitative RT-PCR:

    Article Title: IL-27 mediates HLA class I up-regulation, which can be inhibited by the IL-6 pathway, in HLA-deficient Small Cell Lung Cancer cells
    Article Snippet: .. For immunofluorescence and QRT-PCR analyses, cells were seeded in 24-well plates in culture medium at 5 × 104 cells/well and different cytokines were added: IFN-γ (1000 IU/ml, PeproTech, 300–02), IL-27 (100 ng/ml R & D System, 2526-IL-010), IL-6 (50 ng/ml R & D System 206-IL-010) or recombinant human IL-6Rα/IL-6 chimera [sIL-6R/IL-6] (50 ng/ml R & D System 8954-SR-025). .. Treatments were carried out for 48 h. For the analysis of tyrosine-phosphorylated STAT proteins, 1 × 105 SCLC cells were incubated in a test tube at 37 °C with or without 50 ng/ ml of IL-27, 20 ng/ml of IL-6, 40 ng/ml of sIL-6R/IL-6 in 0.5 ml of medium for the 10, 30 or 60 min time points.

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    R&D Systems il 6 soluble receptor
    Effects of red blood cell (RBC) supernatants (SUP) on LPS or TNF-α-induced inflammatory response of human pulmonary artery endothelial cells (HPAECs). A and B : HPAECs were treated with LPS ( A ) or TNF-α ( B ) for 1 h followed by addition of fresh RBC supernatants (1:100 dilution) for 16 to 20 h and ELISA analysis of the markers of inflammation: soluble ICAM-1 (sICAM-1), IL-8, and <t>IL-6;</t> n = 3. * P
    Il 6 Soluble Receptor, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    R&D Systems recombinant human sgp130 fc
    IL6 trans-signaling converts non-stem cells into stem-like cells. a MCF 10A spheres cultured without or with IL6, IL6 plus <t>sgp130-Fc</t> or with HIL6. b CFSE-labeled MCF 10A cells were cultured as spheres with or without activators (IL6, HIL6) and inhibitors of classical (an anti-IL6 antibody) and trans-signaling (sgp130-Fc). CFSE-dilution in CD44 high CD24 low , CD44 high CD24 high and CD44 low CD24 high/intermediate cells was determined by flow cytometry at day 4. The CFSE-fluorescence intensity of all cells at day one is included as reference. Data are representative for three 3 independently performed experiments. c The absolute number of CD44 high CD24 low , CD44 high CD24 high , CD44 low CD24 high/intermediate cells (upper panel) and LRCs (CFSE high , lower panel) was determined as cell/bead ratio at day 4 by flow cytometry (N=4-5 per group). d Fold-change correlation analysis comparing gene expression changes induced by IL6 plus sgp130 (classical signaling) and HIL6 (trans signaling) in MCF 10 A cells at 12 and 24 hrs with the gene expression signatures of luminal progenitor (LumProg), mature luminal (MatLum) and mammary stem cell enriched cells (MaSC) according to the study of Lim et al. 35 Nc cor: non-centered correlation between fold-changes, Num: number of common differentially expressed genes. e nLRCs from primary, PKH26-labelled control mammosphere-cultures were sorted by flow cytometry as PKH - cells. f Primary HMECs were cultured as spheres for two consecutive rounds in the absence (N=26) or presence of HIL6 and IL6 (HIL6+HIL6, N=18; IL6+HIL6, N=15; HIL6+IL6, N=14; IL6+IL6, N=17). P values in panel c: one-way ANOVA with Dunett’s multiple comparisons test (post-hoc); panel d: P values according to Student’s t-distribution for Nc cor and hypergeometric testing for Num. panel f: one-way ANOVA with Tukey’s multiple comparisons test (post-hoc); comparisons between groups labeled in red are depicted in the bar graph. Asterisks indicate significance between groups (* P
    Recombinant Human Sgp130 Fc, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems soluble gp130
    Human IL-6 engineered with complex N -glycans fails to bind <t>gp130</t> independently of IL-6Rα. ELISA assay testing gp130 binding by wild-type, E109T ( A ) or F102N ( B ) huIL-6 in the presence (+) or absence (−) of sIL-6Rα. Equal amounts
    Soluble Gp130, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    R&D Systems recombinant sgp130
    Inhibition of IL ‐6 trans‐signalling alleviates HG ‐induced podocyte injury. ( A ) Western blotting analysis and ( B ) summarized data presenting the protein expression of p‐ STAT 3 Tyr 705, p‐ STAT 3 Ser 727, STAT 3 and desmin in HG ‐stimulated podocytes pre‐treated without or with recombinant soluble gp130 <t>(sgp130)</t> for 1 hr (p‐ STAT 3 Ser 727 summarized data not presented, P > 0.05). n = 5, * P
    Recombinant Sgp130, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of red blood cell (RBC) supernatants (SUP) on LPS or TNF-α-induced inflammatory response of human pulmonary artery endothelial cells (HPAECs). A and B : HPAECs were treated with LPS ( A ) or TNF-α ( B ) for 1 h followed by addition of fresh RBC supernatants (1:100 dilution) for 16 to 20 h and ELISA analysis of the markers of inflammation: soluble ICAM-1 (sICAM-1), IL-8, and IL-6; n = 3. * P

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

    Article Title: Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

    doi: 10.1152/ajplung.00025.2019

    Figure Lengend Snippet: Effects of red blood cell (RBC) supernatants (SUP) on LPS or TNF-α-induced inflammatory response of human pulmonary artery endothelial cells (HPAECs). A and B : HPAECs were treated with LPS ( A ) or TNF-α ( B ) for 1 h followed by addition of fresh RBC supernatants (1:100 dilution) for 16 to 20 h and ELISA analysis of the markers of inflammation: soluble ICAM-1 (sICAM-1), IL-8, and IL-6; n = 3. * P

    Article Snippet: Human recombinant TNF-α, IL-6, IL-6-soluble receptor, and ELISA kits for human soluble ICAM-1 (sICAM-1)/IL-8 were obtained from R & D Systems (Minneapolis, MN); lipopolysaccharide (LPS; Escherichia coli O55:B5); and antibodies for human ICAM-1/VCAM-1 were from Santa Cruz Biotechnology (Santa Cruz, CA); VE-cadherin antibody was from Cayman Chemical (Ann Arbor, MI); and antibodies for IκBα and β-tubulin were from Cell Signaling (Danvers, MA).

    Techniques: Enzyme-linked Immunosorbent Assay

    Effects of red blood cell (RBC) supernatants (SUP) on inflammatory response by human pulmonary artery endothelial cells (HPAECs). HPAECs were challenged with RBC supernatants from fresh and old stored RBCs at 1:50 and 1:100 dilutions, or cells were stimulated with LPS as a positive control. A – C : after 16–20 h, the cultured medium was examined by ELISA for inflammatory markers: soluble ICAM-1 (sICAM-1) ( A ), IL-8 ( B ), and IL-6 ( C ); n = 5. * P

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

    Article Title: Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

    doi: 10.1152/ajplung.00025.2019

    Figure Lengend Snippet: Effects of red blood cell (RBC) supernatants (SUP) on inflammatory response by human pulmonary artery endothelial cells (HPAECs). HPAECs were challenged with RBC supernatants from fresh and old stored RBCs at 1:50 and 1:100 dilutions, or cells were stimulated with LPS as a positive control. A – C : after 16–20 h, the cultured medium was examined by ELISA for inflammatory markers: soluble ICAM-1 (sICAM-1) ( A ), IL-8 ( B ), and IL-6 ( C ); n = 5. * P

    Article Snippet: Human recombinant TNF-α, IL-6, IL-6-soluble receptor, and ELISA kits for human soluble ICAM-1 (sICAM-1)/IL-8 were obtained from R & D Systems (Minneapolis, MN); lipopolysaccharide (LPS; Escherichia coli O55:B5); and antibodies for human ICAM-1/VCAM-1 were from Santa Cruz Biotechnology (Santa Cruz, CA); VE-cadherin antibody was from Cayman Chemical (Ann Arbor, MI); and antibodies for IκBα and β-tubulin were from Cell Signaling (Danvers, MA).

    Techniques: Positive Control, Cell Culture, Enzyme-linked Immunosorbent Assay

    IL6 trans-signaling converts non-stem cells into stem-like cells. a MCF 10A spheres cultured without or with IL6, IL6 plus sgp130-Fc or with HIL6. b CFSE-labeled MCF 10A cells were cultured as spheres with or without activators (IL6, HIL6) and inhibitors of classical (an anti-IL6 antibody) and trans-signaling (sgp130-Fc). CFSE-dilution in CD44 high CD24 low , CD44 high CD24 high and CD44 low CD24 high/intermediate cells was determined by flow cytometry at day 4. The CFSE-fluorescence intensity of all cells at day one is included as reference. Data are representative for three 3 independently performed experiments. c The absolute number of CD44 high CD24 low , CD44 high CD24 high , CD44 low CD24 high/intermediate cells (upper panel) and LRCs (CFSE high , lower panel) was determined as cell/bead ratio at day 4 by flow cytometry (N=4-5 per group). d Fold-change correlation analysis comparing gene expression changes induced by IL6 plus sgp130 (classical signaling) and HIL6 (trans signaling) in MCF 10 A cells at 12 and 24 hrs with the gene expression signatures of luminal progenitor (LumProg), mature luminal (MatLum) and mammary stem cell enriched cells (MaSC) according to the study of Lim et al. 35 Nc cor: non-centered correlation between fold-changes, Num: number of common differentially expressed genes. e nLRCs from primary, PKH26-labelled control mammosphere-cultures were sorted by flow cytometry as PKH - cells. f Primary HMECs were cultured as spheres for two consecutive rounds in the absence (N=26) or presence of HIL6 and IL6 (HIL6+HIL6, N=18; IL6+HIL6, N=15; HIL6+IL6, N=14; IL6+IL6, N=17). P values in panel c: one-way ANOVA with Dunett’s multiple comparisons test (post-hoc); panel d: P values according to Student’s t-distribution for Nc cor and hypergeometric testing for Num. panel f: one-way ANOVA with Tukey’s multiple comparisons test (post-hoc); comparisons between groups labeled in red are depicted in the bar graph. Asterisks indicate significance between groups (* P

    Journal: bioRxiv

    Article Title: Interleukin-6 trans-signaling is a candidate mechanism to drive progression of human DCCs during periods of clinical latency

    doi: 10.1101/2020.05.28.121145

    Figure Lengend Snippet: IL6 trans-signaling converts non-stem cells into stem-like cells. a MCF 10A spheres cultured without or with IL6, IL6 plus sgp130-Fc or with HIL6. b CFSE-labeled MCF 10A cells were cultured as spheres with or without activators (IL6, HIL6) and inhibitors of classical (an anti-IL6 antibody) and trans-signaling (sgp130-Fc). CFSE-dilution in CD44 high CD24 low , CD44 high CD24 high and CD44 low CD24 high/intermediate cells was determined by flow cytometry at day 4. The CFSE-fluorescence intensity of all cells at day one is included as reference. Data are representative for three 3 independently performed experiments. c The absolute number of CD44 high CD24 low , CD44 high CD24 high , CD44 low CD24 high/intermediate cells (upper panel) and LRCs (CFSE high , lower panel) was determined as cell/bead ratio at day 4 by flow cytometry (N=4-5 per group). d Fold-change correlation analysis comparing gene expression changes induced by IL6 plus sgp130 (classical signaling) and HIL6 (trans signaling) in MCF 10 A cells at 12 and 24 hrs with the gene expression signatures of luminal progenitor (LumProg), mature luminal (MatLum) and mammary stem cell enriched cells (MaSC) according to the study of Lim et al. 35 Nc cor: non-centered correlation between fold-changes, Num: number of common differentially expressed genes. e nLRCs from primary, PKH26-labelled control mammosphere-cultures were sorted by flow cytometry as PKH - cells. f Primary HMECs were cultured as spheres for two consecutive rounds in the absence (N=26) or presence of HIL6 and IL6 (HIL6+HIL6, N=18; IL6+HIL6, N=15; HIL6+IL6, N=14; IL6+IL6, N=17). P values in panel c: one-way ANOVA with Dunett’s multiple comparisons test (post-hoc); panel d: P values according to Student’s t-distribution for Nc cor and hypergeometric testing for Num. panel f: one-way ANOVA with Tukey’s multiple comparisons test (post-hoc); comparisons between groups labeled in red are depicted in the bar graph. Asterisks indicate significance between groups (* P

    Article Snippet: mRNA microarray experimentsMCF 10A cells were cultured as mammospheres in the presence or absence of 10 ng/ml IL6 (Sigma-Aldrich, Germany), 10 ng/ml IL6 + 0.1 ng/ml recombinant human sgp130-Fc (R & D Systems, Germany) or 20 ng/ml Hyper-IL6 (kind gift of S. Rose-John, Christian-Albrechts-University, Germany) for 12 and 24 hours.

    Techniques: Cell Culture, Labeling, Flow Cytometry, Fluorescence, Expressing

    IL6 trans-signaling regulates the frequency of MCF 10A, hTERT-HME1 and primary HMECs with sphere-forming ability. a MCF 10A cells were cultured as spheres in the absence (N=18) or presence of IL6 (N=18), an IL6-blocking antibody (N=6) or Hyper-IL6 (N=6). b hTERT-HME1 were cultured as spheres in the absence (N=3) or presence (N=3) of Hyper-IL6. c HMECs were cultured without or with IL6, with an IL6 blocking antibody or Hyper-IL6. N=3 patients, each patient analyzed in triplicate. d hTERT-HME1-EGFR Δ746–750 cells were cultured as spheres in the absence or presence of HIL6 (each N=12). e MCF 10A cells were cultured as spheres without (N=6) or with IL6 (N=6) and IL6 plus sgp130-Fc at indicated concentrations (each N=6). f Sphere formation of hTERT-HME1-EGFR Δ746-750 in the absence (N=10) or presence of an anti-IL6 antibody (N=9) or with sgp130-Fc at indicated concentrations (each N=12). Cumulative data of three experiments. P values in panel a, c, f: one-way ANOVA with Dunnett’s multiple comparisons test (post hoc); panel b, d: two-sided Student’s t-test; panel e: one-way ANOVA with Tukey’s multiple comparisons test (post hoc); asterisks indicate significance between groups (*P

    Journal: bioRxiv

    Article Title: Interleukin-6 trans-signaling is a candidate mechanism to drive progression of human DCCs during periods of clinical latency

    doi: 10.1101/2020.05.28.121145

    Figure Lengend Snippet: IL6 trans-signaling regulates the frequency of MCF 10A, hTERT-HME1 and primary HMECs with sphere-forming ability. a MCF 10A cells were cultured as spheres in the absence (N=18) or presence of IL6 (N=18), an IL6-blocking antibody (N=6) or Hyper-IL6 (N=6). b hTERT-HME1 were cultured as spheres in the absence (N=3) or presence (N=3) of Hyper-IL6. c HMECs were cultured without or with IL6, with an IL6 blocking antibody or Hyper-IL6. N=3 patients, each patient analyzed in triplicate. d hTERT-HME1-EGFR Δ746–750 cells were cultured as spheres in the absence or presence of HIL6 (each N=12). e MCF 10A cells were cultured as spheres without (N=6) or with IL6 (N=6) and IL6 plus sgp130-Fc at indicated concentrations (each N=6). f Sphere formation of hTERT-HME1-EGFR Δ746-750 in the absence (N=10) or presence of an anti-IL6 antibody (N=9) or with sgp130-Fc at indicated concentrations (each N=12). Cumulative data of three experiments. P values in panel a, c, f: one-way ANOVA with Dunnett’s multiple comparisons test (post hoc); panel b, d: two-sided Student’s t-test; panel e: one-way ANOVA with Tukey’s multiple comparisons test (post hoc); asterisks indicate significance between groups (*P

    Article Snippet: mRNA microarray experimentsMCF 10A cells were cultured as mammospheres in the presence or absence of 10 ng/ml IL6 (Sigma-Aldrich, Germany), 10 ng/ml IL6 + 0.1 ng/ml recombinant human sgp130-Fc (R & D Systems, Germany) or 20 ng/ml Hyper-IL6 (kind gift of S. Rose-John, Christian-Albrechts-University, Germany) for 12 and 24 hours.

    Techniques: Cell Culture, Blocking Assay

    Human IL-6 engineered with complex N -glycans fails to bind gp130 independently of IL-6Rα. ELISA assay testing gp130 binding by wild-type, E109T ( A ) or F102N ( B ) huIL-6 in the presence (+) or absence (−) of sIL-6Rα. Equal amounts

    Journal:

    Article Title: Complex N-Linked Glycans on Asn-89 of Kaposi Sarcoma Herpes Virus-encoded Interleukin-6 Mediate Optimal Function by Affecting Cytokine Protein Conformation *

    doi: 10.1074/jbc.M109.039115

    Figure Lengend Snippet: Human IL-6 engineered with complex N -glycans fails to bind gp130 independently of IL-6Rα. ELISA assay testing gp130 binding by wild-type, E109T ( A ) or F102N ( B ) huIL-6 in the presence (+) or absence (−) of sIL-6Rα. Equal amounts

    Article Snippet: ELISA plates were coated with soluble gp130 (R & D Systems) at 625 ng/ml in carbonate buffer overnight.

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

    A model for the differing mechanism of action of huIL-6 and vIL-6. A , binding of human IL-6 to IL-6Rα changes the conformation of IL-6 to allow for binding to gp130. B , the complex N -linked glycan on Asn-89 of vIL-6 leads to a conformational alteration,

    Journal:

    Article Title: Complex N-Linked Glycans on Asn-89 of Kaposi Sarcoma Herpes Virus-encoded Interleukin-6 Mediate Optimal Function by Affecting Cytokine Protein Conformation *

    doi: 10.1074/jbc.M109.039115

    Figure Lengend Snippet: A model for the differing mechanism of action of huIL-6 and vIL-6. A , binding of human IL-6 to IL-6Rα changes the conformation of IL-6 to allow for binding to gp130. B , the complex N -linked glycan on Asn-89 of vIL-6 leads to a conformational alteration,

    Article Snippet: ELISA plates were coated with soluble gp130 (R & D Systems) at 625 ng/ml in carbonate buffer overnight.

    Techniques: Binding Assay

    Inhibition of IL ‐6 trans‐signalling alleviates HG ‐induced podocyte injury. ( A ) Western blotting analysis and ( B ) summarized data presenting the protein expression of p‐ STAT 3 Tyr 705, p‐ STAT 3 Ser 727, STAT 3 and desmin in HG ‐stimulated podocytes pre‐treated without or with recombinant soluble gp130 (sgp130) for 1 hr (p‐ STAT 3 Ser 727 summarized data not presented, P > 0.05). n = 5, * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: The classic signalling and trans‐signalling of interleukin‐6 are both injurious in podocyte under high glucose exposure

    doi: 10.1111/jcmm.13314

    Figure Lengend Snippet: Inhibition of IL ‐6 trans‐signalling alleviates HG ‐induced podocyte injury. ( A ) Western blotting analysis and ( B ) summarized data presenting the protein expression of p‐ STAT 3 Tyr 705, p‐ STAT 3 Ser 727, STAT 3 and desmin in HG ‐stimulated podocytes pre‐treated without or with recombinant soluble gp130 (sgp130) for 1 hr (p‐ STAT 3 Ser 727 summarized data not presented, P > 0.05). n = 5, * P

    Article Snippet: After individual pre‐treatment with gp130 antibody (2 μg/ml, R & D Systems, Minneapolis, MN, USA), IL‐6 antibody (1 μg/ml, R & D Systems, Minneapolis, MN, USA), recombinant sgp130 (1 μg/ml, R & D Systems, Minneapolis, MN, USA), recombinant human IL‐6 (20 ng/ml, Peprotech, Rocky Hill, NJ, USA) or complex of IL‐6 and recombinant human soluble IL‐6R protein (30 ng/ml, R & D Systems, Minneapolis, MN, USA), podocytes were exposed to HG or osmotic control for 24 hrs.

    Techniques: Inhibition, Western Blot, Expressing, Recombinant

    Circulatory IL ‐6, sIL ‐6R and sgp130 abundance is increased in DKD patients. ELISA analysis data showing IL ‐6 ( A ), sIL ‐6R ( B ) and sgp130 ( C ) levels in serum samples from healthy controls (Ctrl) and DKD patients ( DKD ). n = 10, * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: The classic signalling and trans‐signalling of interleukin‐6 are both injurious in podocyte under high glucose exposure

    doi: 10.1111/jcmm.13314

    Figure Lengend Snippet: Circulatory IL ‐6, sIL ‐6R and sgp130 abundance is increased in DKD patients. ELISA analysis data showing IL ‐6 ( A ), sIL ‐6R ( B ) and sgp130 ( C ) levels in serum samples from healthy controls (Ctrl) and DKD patients ( DKD ). n = 10, * P

    Article Snippet: After individual pre‐treatment with gp130 antibody (2 μg/ml, R & D Systems, Minneapolis, MN, USA), IL‐6 antibody (1 μg/ml, R & D Systems, Minneapolis, MN, USA), recombinant sgp130 (1 μg/ml, R & D Systems, Minneapolis, MN, USA), recombinant human IL‐6 (20 ng/ml, Peprotech, Rocky Hill, NJ, USA) or complex of IL‐6 and recombinant human soluble IL‐6R protein (30 ng/ml, R & D Systems, Minneapolis, MN, USA), podocytes were exposed to HG or osmotic control for 24 hrs.

    Techniques: Enzyme-linked Immunosorbent Assay