soluble il 6rα  (R&D Systems)

 
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    Name:
    Recombinant Human IL 6 R alpha Protein CF
    Description:
    The Recombinant Human IL 6 R alpha Protein from R D Systems is derived from Sf 21 baculovirus The Recombinant Human IL 6 R alpha Protein has been validated for the following applications Bioactivity
    Catalog Number:
    227-SR-025/CF
    Price:
    379
    Category:
    Proteins and Enzymes
    Source:
    Sf 21 (baculovirus)-derived Recombinant Human IL-6 R alpha Protein
    Applications:
    Bioactivity
    Purity:
    >97%, by SDS-PAGE under reducing conditions and visualized by silver stain
    Conjugate:
    Unconjugated
    Size:
    25 ug
    Buy from Supplier


    Structured Review

    R&D Systems soluble il 6rα
    Recombinant Human IL 6 R alpha Protein CF
    The Recombinant Human IL 6 R alpha Protein from R D Systems is derived from Sf 21 baculovirus The Recombinant Human IL 6 R alpha Protein has been validated for the following applications Bioactivity
    https://www.bioz.com/result/soluble il 6rα/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    soluble il 6rα - by Bioz Stars, 2021-06
    93/100 stars

    Images

    1) Product Images from "Investigation of fascin1, a marker of mature dendritic cells, reveals a New role for IL-6 signaling in chemotaxis"

    Article Title: Investigation of fascin1, a marker of mature dendritic cells, reveals a New role for IL-6 signaling in chemotaxis

    Journal: bioRxiv

    doi: 10.1101/2020.03.19.979104

    IL-6 signaling is critical for chemotaxis of DCs. A , Higher IL-6 signaling of WT BMDCs than that of fascin1 KO BMDCs. Proximity Ligation Assay (PLA) was used to determine the in situ association between IL-6Rα and gp130, representing the extent of IL-6 signaling. a b, immature BMDCs; c d, mature BMDCs. a c, WT. b d, fascin1 KO. Fluorescence speckles (arrows) indicate the association between IL-6Rα and gp130. Cell boundaries are indicated by dashed lines. B , Quantitative analyses of PLA signals in WT and fascin1 KO BMDCs (n=51). **, p
    Figure Legend Snippet: IL-6 signaling is critical for chemotaxis of DCs. A , Higher IL-6 signaling of WT BMDCs than that of fascin1 KO BMDCs. Proximity Ligation Assay (PLA) was used to determine the in situ association between IL-6Rα and gp130, representing the extent of IL-6 signaling. a b, immature BMDCs; c d, mature BMDCs. a c, WT. b d, fascin1 KO. Fluorescence speckles (arrows) indicate the association between IL-6Rα and gp130. Cell boundaries are indicated by dashed lines. B , Quantitative analyses of PLA signals in WT and fascin1 KO BMDCs (n=51). **, p

    Techniques Used: Chemotaxis Assay, Proximity Ligation Assay, In Situ, Fluorescence

    IL-6Rα signaling is critical for CCR7 internalization and CCL19-induced ERK1/2 phosphorylation. A , Effects of the blockage of IL-6 signaling on CCR7 internalization. WT BMDCs were stimulated with CCL19 for 5 min in the presence of either control (a) or neutralizing antibody (b), and then surface CCR7 expression was determined by flow cytometry. Surface expression of CCR7 was examined with CD86 + BMDCs before (-CCL19) and after CCL19 addition (+CCL19). Note that CCR7 internalization was blocked in the absence of a neutralizing antibody to IL-6Rα. B , Effects of IL-6 signaling on CCR7 internalization of IL-6Rα KO BMDCs. IL-6Rα KO BMDCs were stimulated with CCL19 in the absence (a) or presence of sIL-6Rα (soluble IL-6Rα) and IL-6 (b). Note that IL-6Rα KO BMDCs showed impaired CCR7 internalization (a), which was rescued by the addition of sIL-6Rα and IL-6 (b). C , Inhibition of CCR7 internalization by the blockage of IL-6 signaling in HEK293T epithelial cells. HEK293T cells stably expressing mouse CCR7-GFP were stimulated with CCL19 in the presence of either control or neutralizing antibody against IL-6Rα. a-d, fluorescence images of CCR7-GFP in control cells (a b) or IL-6Rα neutralizing antibody-treated cells (c d) before (a c) or after addition of CCL19 (b d). The same cells were counter stained with an antibody specific to mouse CCR7 to detect only surface CCR7 (e-h) in control (e f) or IL-6Rα neutralizing antibody-treated cells (g h) before (e g) or after (f h) addition of CCL19. D , Quantitative measurements of surface CCR7 immunofluorescence (panels e-h of C) at cell-to-cell contacts. In contrast to the control, the addition of a neutralizing antibody against IL-6Rα blocked internalization of CCR7. E , Effects of the blockage of IL-6 signaling on CCL19-induced ERK1/2 phosphorylation of WT BMDCs. WT BMDCs were stimulated with CCL19 in the presence of a control or neutralizing antibody, then ERK1/2 phosphorylation levels were determined at 0, 5, 15 and 30min after CCL19 addition using Western blotting with a phosphospecific ERK1/2 antibody. For normalization, the same membranes were reblotted with a pan ERK1/2 antibody. The ratios of phosphoERK1/2 to total ERK1/2 are indicated below the figure.
    Figure Legend Snippet: IL-6Rα signaling is critical for CCR7 internalization and CCL19-induced ERK1/2 phosphorylation. A , Effects of the blockage of IL-6 signaling on CCR7 internalization. WT BMDCs were stimulated with CCL19 for 5 min in the presence of either control (a) or neutralizing antibody (b), and then surface CCR7 expression was determined by flow cytometry. Surface expression of CCR7 was examined with CD86 + BMDCs before (-CCL19) and after CCL19 addition (+CCL19). Note that CCR7 internalization was blocked in the absence of a neutralizing antibody to IL-6Rα. B , Effects of IL-6 signaling on CCR7 internalization of IL-6Rα KO BMDCs. IL-6Rα KO BMDCs were stimulated with CCL19 in the absence (a) or presence of sIL-6Rα (soluble IL-6Rα) and IL-6 (b). Note that IL-6Rα KO BMDCs showed impaired CCR7 internalization (a), which was rescued by the addition of sIL-6Rα and IL-6 (b). C , Inhibition of CCR7 internalization by the blockage of IL-6 signaling in HEK293T epithelial cells. HEK293T cells stably expressing mouse CCR7-GFP were stimulated with CCL19 in the presence of either control or neutralizing antibody against IL-6Rα. a-d, fluorescence images of CCR7-GFP in control cells (a b) or IL-6Rα neutralizing antibody-treated cells (c d) before (a c) or after addition of CCL19 (b d). The same cells were counter stained with an antibody specific to mouse CCR7 to detect only surface CCR7 (e-h) in control (e f) or IL-6Rα neutralizing antibody-treated cells (g h) before (e g) or after (f h) addition of CCL19. D , Quantitative measurements of surface CCR7 immunofluorescence (panels e-h of C) at cell-to-cell contacts. In contrast to the control, the addition of a neutralizing antibody against IL-6Rα blocked internalization of CCR7. E , Effects of the blockage of IL-6 signaling on CCL19-induced ERK1/2 phosphorylation of WT BMDCs. WT BMDCs were stimulated with CCL19 in the presence of a control or neutralizing antibody, then ERK1/2 phosphorylation levels were determined at 0, 5, 15 and 30min after CCL19 addition using Western blotting with a phosphospecific ERK1/2 antibody. For normalization, the same membranes were reblotted with a pan ERK1/2 antibody. The ratios of phosphoERK1/2 to total ERK1/2 are indicated below the figure.

    Techniques Used: Expressing, Flow Cytometry, Inhibition, Stable Transfection, Fluorescence, Staining, Immunofluorescence, Western Blot

    IL-6 signaling is required for directional migration of BMDCs. A, Analyses of migration tracks. Live cell imaging of BMDCs migrating in a collagen gel toward CCL19 was performed to obtain migration tracks (a, c e) and rose diagram plots for directionality(b, d f). a b, WT (n=19): c d, fascin1 KO (n=23): e f, WT BMDCs in the presence of a neutralizing antibody against IL-6Rα (n=28). WT BMDCs (a b) showed more consistent migration toward CCL19 than fascin1 KO counterparts (c d). Blockage of IL-6 signaling inhibits directed migration of WT BMDCs (e f). Arrows, the direction of a CCL19 gradient. B-E, Box plot analyses of parallel (B, FMI∥) and perpendicular (C, FMI⊥) forward migration indexes, directness (D), and migration speeds (E). *, p
    Figure Legend Snippet: IL-6 signaling is required for directional migration of BMDCs. A, Analyses of migration tracks. Live cell imaging of BMDCs migrating in a collagen gel toward CCL19 was performed to obtain migration tracks (a, c e) and rose diagram plots for directionality(b, d f). a b, WT (n=19): c d, fascin1 KO (n=23): e f, WT BMDCs in the presence of a neutralizing antibody against IL-6Rα (n=28). WT BMDCs (a b) showed more consistent migration toward CCL19 than fascin1 KO counterparts (c d). Blockage of IL-6 signaling inhibits directed migration of WT BMDCs (e f). Arrows, the direction of a CCL19 gradient. B-E, Box plot analyses of parallel (B, FMI∥) and perpendicular (C, FMI⊥) forward migration indexes, directness (D), and migration speeds (E). *, p

    Techniques Used: Migration, Live Cell Imaging

    Impaired chemotaxis of IL-6Rα KO BMDCs and its restoration by the addition of soluble IL-6Rα (sIL-6Rα) and IL-6. BMDCs were isolated from heterozygous (hetero) and IL-6Rα KO mice. A , Immunofluorescence of IL-6Rα in heterozygous (a) and IL-6Rα KO BMDCs (b) confirming the lack of IL-6Rα in KO BMDCs. B , Collagen-coated, modified Boyden chamber chemotaxis assays of heterozygous (hetero), IL-6Rα KO, and IL-6Rα KO in the presence of sIL-6Rα and IL-6 (KO+sIL-6R/IL-6). Note that KO BMDCs show reduced chemotaxis, which was rescued by the addition of soluble IL-6Rα (sIL-6Rα) and IL-6. C D , IL-6Rα KO did not alter surface expression of CCR7. Immunofluorescence (C) showing similar levels of surface CCR7 in heterozygous (a) and IL-6Rα KO BMDCs (b). Flow cytometry analyses (D) confirming that IL-6Rα KO did not affect the levels of CCR7 surface expression. After CD86-positive heterozygous (a) and IL-6Rα KO (b) BMDCs were gated, the levels of CCR7 surface expression were examined in histogram (c).
    Figure Legend Snippet: Impaired chemotaxis of IL-6Rα KO BMDCs and its restoration by the addition of soluble IL-6Rα (sIL-6Rα) and IL-6. BMDCs were isolated from heterozygous (hetero) and IL-6Rα KO mice. A , Immunofluorescence of IL-6Rα in heterozygous (a) and IL-6Rα KO BMDCs (b) confirming the lack of IL-6Rα in KO BMDCs. B , Collagen-coated, modified Boyden chamber chemotaxis assays of heterozygous (hetero), IL-6Rα KO, and IL-6Rα KO in the presence of sIL-6Rα and IL-6 (KO+sIL-6R/IL-6). Note that KO BMDCs show reduced chemotaxis, which was rescued by the addition of soluble IL-6Rα (sIL-6Rα) and IL-6. C D , IL-6Rα KO did not alter surface expression of CCR7. Immunofluorescence (C) showing similar levels of surface CCR7 in heterozygous (a) and IL-6Rα KO BMDCs (b). Flow cytometry analyses (D) confirming that IL-6Rα KO did not affect the levels of CCR7 surface expression. After CD86-positive heterozygous (a) and IL-6Rα KO (b) BMDCs were gated, the levels of CCR7 surface expression were examined in histogram (c).

    Techniques Used: Chemotaxis Assay, Isolation, Mouse Assay, Immunofluorescence, Modification, Expressing, Flow Cytometry

    2) Product Images from "A mutual activation loop between breast cancer cells and myeloid-derived suppressor cells facilitates spontaneous metastasis through IL-6 trans-signaling in a murine model"

    Article Title: A mutual activation loop between breast cancer cells and myeloid-derived suppressor cells facilitates spontaneous metastasis through IL-6 trans-signaling in a murine model

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/bcr3473

    Increased shedding of sIL-6Rα by the MDSCs of tumor-bearing mice . (A) Soluble IL-6Rα levels in culture supernatants of splenic MDSCs were measured by ELISA and (B) surface IL-6Rα levels on splenic MDSCs were measured by FACS. (C) The mRNA expression of Adam10 and Adam17 in splenic MDSCs of naïve and tumor-bearing mice were determined by qRT-PCR. (D-E) Protease inhibitor cocktails were applied to cultures of splenic MDSCs from 4T1 cell-bearing mice for 6 or 24 hours. (D) Membrane-bound IL-6Rα was detected by FACS and (E) soluble IL-6Rα levels were measured by ELISA. (F) Tissue sections were stained for Adam17 (green), Gr-1 (red), and DAPI (blue) to compare their localizations. Scale bar = 30 μm (original magnification, ×1,000). FACS, fluorescence-activated cell sorting; MDSCs, myeloid-derived suppressor cells.
    Figure Legend Snippet: Increased shedding of sIL-6Rα by the MDSCs of tumor-bearing mice . (A) Soluble IL-6Rα levels in culture supernatants of splenic MDSCs were measured by ELISA and (B) surface IL-6Rα levels on splenic MDSCs were measured by FACS. (C) The mRNA expression of Adam10 and Adam17 in splenic MDSCs of naïve and tumor-bearing mice were determined by qRT-PCR. (D-E) Protease inhibitor cocktails were applied to cultures of splenic MDSCs from 4T1 cell-bearing mice for 6 or 24 hours. (D) Membrane-bound IL-6Rα was detected by FACS and (E) soluble IL-6Rα levels were measured by ELISA. (F) Tissue sections were stained for Adam17 (green), Gr-1 (red), and DAPI (blue) to compare their localizations. Scale bar = 30 μm (original magnification, ×1,000). FACS, fluorescence-activated cell sorting; MDSCs, myeloid-derived suppressor cells.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, FACS, Expressing, Quantitative RT-PCR, Protease Inhibitor, Staining, Fluorescence, Derivative Assay

    Activated MDSCs contributed to tumor invasiveness through IL-6 trans-signaling . (A-B) 4T1 cells were treated with recombinant IL-6 plus soluble IL-6Rα (A) or 4T1/MDSC-CM (B) for 30 minutes in the presence of anti-IL-6R blocking antibody or gp130-Fc. (C) 4T1 cells were allowed to invade through Matrigel for 18 hours in the presence or absence of 4T1/MDSC-CM and/or gp130-Fc (crystal violet). (D-E) 4T1 cells were injected into the mammary fat pads. Some mice underwent continuous administration using osmotic mini-pumps (5 or 10 μg for 14 days). (D) Primary tumor growth and (E) numbers of metastatic masses in the lungs at 26 days. (F) 4T1 cells were injected intravenously into BALB/c mice ( n = 5 mice per group). Some mice received gp130-Fc (2.5 μg) 4 days after cancer cell injection. Numbers of metastatic masses in the lungs at day 12 were determined. Values are the means ± SEM of each group. * P
    Figure Legend Snippet: Activated MDSCs contributed to tumor invasiveness through IL-6 trans-signaling . (A-B) 4T1 cells were treated with recombinant IL-6 plus soluble IL-6Rα (A) or 4T1/MDSC-CM (B) for 30 minutes in the presence of anti-IL-6R blocking antibody or gp130-Fc. (C) 4T1 cells were allowed to invade through Matrigel for 18 hours in the presence or absence of 4T1/MDSC-CM and/or gp130-Fc (crystal violet). (D-E) 4T1 cells were injected into the mammary fat pads. Some mice underwent continuous administration using osmotic mini-pumps (5 or 10 μg for 14 days). (D) Primary tumor growth and (E) numbers of metastatic masses in the lungs at 26 days. (F) 4T1 cells were injected intravenously into BALB/c mice ( n = 5 mice per group). Some mice received gp130-Fc (2.5 μg) 4 days after cancer cell injection. Numbers of metastatic masses in the lungs at day 12 were determined. Values are the means ± SEM of each group. * P

    Techniques Used: Recombinant, Blocking Assay, Injection, Mouse Assay

    Related Articles

    Recombinant:

    Article Title: Orthogonal Mass Spectrometry-Based Footprinting for Epitope Mapping and Structural Characterization: The IL-6 Receptor upon Binding of Protein Therapeutics
    Article Snippet: Although an exhaustive evaluation by one approach will often provide an answer, we have chosen an integrative course, and the results presented here demonstrate the value of applying that approach for epitope mapping and HOS characterization. .. Recombinant human IL-6R alpha extracellular region (residue 20-358, referred as IL-6R below) was purchased from R & D systems (Minneapolis, MN). .. Adnectin1 ( Kd ∼ 6.2 pM) and adnectin2 ( Kd ∼ 46 nM) were expressed and purified at BMS as previously described.

    Article Title: A mutual activation loop between breast cancer cells and myeloid-derived suppressor cells facilitates spontaneous metastasis through IL-6 trans-signaling in a murine model
    Article Snippet: For IL-6 detection, anti-mouse IL-6 (eBioscience) was used as the capture antibody, biotinylated anti-mouse IL-6 (eBioscience) in 0.1% BSA in PBS/T as the detection antibody and recombinant IL-6 (eBioscience) as the standard. .. To detect soluble IL-6Rα, we used anti-mouse IL-6Rα (R & D Systems) as the capture antibody, biotinylated anti-mouse IL-6Rα (R & D Systems) as the detection antibody and recombinant IL-6Rα (R & D Systems) as the standard. .. RNA analysis RNA was isolated from sorted splenic MDSC using the RNeasy kit (QIAGEN; 74104, Hilden, Germany). cDNA was generated from 1 μg of total RNA by reverse transcriptase from Moloney Murine Leukemia Virus (M-MLV) (TAKARA, Shiga, Japan), and subjected to PCR.

    Article Title: Regulation of TRPM7 Function by IL-6 through the JAK2-STAT3 Signaling Pathway
    Article Snippet: .. Compounds Pharmacological compounds used in this study included recombinant human IL-6 (Preprotech, London, UK), recombinant human IL-6R (Preprotech), rabbit monoclonal antibody against IL-6R (R & D Systems, Inc. Minneapolis, MN, USA), and goat polyclonal antibody against TRPM7 (Santa Cruz Biotechnology, Santa Cruz, CA, USA). .. The following compounds were purchased from Sigma−Aldrich (St. Louis, MO, USA): the JAK2 inhibitor AG490, the PI3K inhibitor wortmannin, the PLC inhibitor U73122, TEACL, TTX, nimodipine, CsCl, choline chloride, spermine, GdCl3, and 2-APB.

    Article Title: Functional Heterogeneity of Breast Fibroblasts Is Defined by a Prostaglandin Secretory Phenotype that Promotes Expansion of Cancer-Stem Like Cells
    Article Snippet: For MCF7 treatments, cells were grown in PRF-DMEM supplemented with 5% charcoal/dextran stripped FBS, 1% AB/AM, and treated with ethanol, 0.5 µM PGE2, or 1 nM 17-β-Estradiol (Sigma Aldrich). .. For MCF7 treatments involving fibroblast CM, CM was collected as previously described, supplemented with 5% charcoal/dextran stripped FBS, and administered to MCF7 cells for 6 days. α-IL6 and recombinant human IL-6 (R & D Systems, Minneapolis, MN) were used at 1.5 µg/mL and 10 ng/mL, respectively. .. Preparation of cells for mouse mammary fat pad inoculation All animal procedures were performed in accordance with an approved protocol by Tufts University Institutional Animal Care and Use Committee.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Activation of STAT3 signaling is mediated by TFF1 silencing in gastric neoplasia
    Article Snippet: Based on the PLA results and these findings, it is more likely that TFF1 binds to IL6Rα to interfere with binding of the ligand, IL6. .. To confirm that TFF1 can indeed interfere with IL6–IL6Rα, we performed a quantitative ELISA assay (R & D systems) and checked the levels of soluble IL6–IL6Rα complex formation, after stimulation with IL6. .. AGS cells were infected with TFF1 or control adenoviruses.

    Article Title: Differential baseline and response profile to IFN-? gene transduction of IL-6/IL-6 receptor-? secretion discriminate primary tumors versus bone marrow metastases of nasopharyngeal carcinomas in culture
    Article Snippet: Appropriate positive and negative control antibodies, such as anti-HLA class I (mAb W6/32), phosphate-buffered saline (PBS) and isotype-matched irrelevant mAbs were included in each experiment. .. Enzyme-linked immunosorbent assay (ELISA) ELISA kits for quantification of human IFN-γ, IL-6 and IL-6Rα were all purchased from R & D Systems, Inc., Minneapolis, MN. ..

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    R&D Systems sil 6r
    Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor <t>(sIL-6R)</t> expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
    Sil 6r, 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
    https://www.bioz.com/result/sil 6r/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sil 6r - by Bioz Stars, 2021-06
    93/100 stars
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    Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

    Journal: Carcinogenesis

    Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

    doi: 10.1093/carcin/bgr075

    Figure Lengend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

    Article Snippet: Most importantly, blocking sIL-6R with sIL-6R-specific neutralizing antibody completely abolished this growth-promoting effect ( P < 0.01, ) but a matched isotype control antibody could not block the effect.

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

    Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

    Journal: Carcinogenesis

    Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

    doi: 10.1093/carcin/bgr075

    Figure Lengend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

    Article Snippet: Most importantly, blocking sIL-6R with sIL-6R-specific neutralizing antibody completely abolished this growth-promoting effect ( P < 0.01, ) but a matched isotype control antibody could not block the effect.

    Techniques: Blocking Assay, Expressing, MTT Assay

    Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

    Journal: Journal of neuroimmunology

    Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

    doi: 10.1016/j.jneuroim.2018.09.002

    Figure Lengend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

    Article Snippet: The cerebrospinal fluid of aged mice had increased amounts of sIL-6R compared with adult counterparts, although adult and aged mice did not produce significantly different amounts of its inhibitor, sgp130 ( ).

    Techniques: Enzyme-linked Immunosorbent Assay

    Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

    Journal: Journal of neuroimmunology

    Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

    doi: 10.1016/j.jneuroim.2018.09.002

    Figure Lengend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

    Article Snippet: The cerebrospinal fluid of aged mice had increased amounts of sIL-6R compared with adult counterparts, although adult and aged mice did not produce significantly different amounts of its inhibitor, sgp130 ( ).

    Techniques: Expressing

    sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

    Journal: Journal of neuroimmunology

    Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

    doi: 10.1016/j.jneuroim.2018.09.002

    Figure Lengend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

    Article Snippet: The cerebrospinal fluid of aged mice had increased amounts of sIL-6R compared with adult counterparts, although adult and aged mice did not produce significantly different amounts of its inhibitor, sgp130 ( ).

    Techniques: Expressing, Dot Blot, Fluorescence

    IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P

    Journal: Journal of Neuroinflammation

    Article Title: Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior

    doi: 10.1186/1742-2094-8-54

    Figure Lengend Snippet: IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P

    Article Snippet: Here, STAT3 was upregulated in response to both IL-6 and LPS in BV.2 and Neuro.2A cells and pretreatment with sIL-6R led to an increased IL-6- and LPS-induced STAT3 phosphorylation.

    Techniques: