sil 6r alpha  (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
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    Structured Review

    R&D Systems sil 6r alpha
    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/sil 6r alpha/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sil 6r alpha - by Bioz Stars, 2021-05
    93/100 stars

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    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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  • 93
    R&D Systems soluble il 6rα
    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 <t>IL-6Rα</t> 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
    Soluble Il 6rα, 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/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, 2021-05
    93/100 stars
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    95
    R&D Systems sil 6rα
    Inflammatory profile of liver and EWAT (8-week HFD). ( a ) Liver ( n =10 versus 12) and ( b ) EWAT ( n =11) gene expression profile. Activity of c-Jun kinase (JNK) in ( c ) liver and ( d ) EWAT with the measurement of pS63 c-Jun by immunoblotting (representative immunoblots of 2 blots) and quantification ( n =8); relative unit (RU) to that of <t>IL-6Rα</t> f/f (Control). Flow cytometry analyses of ( e ) liver and ( f ) EWAT composition of total T cells, CD8+ and CD4+ T cells, T helper cells (Th1, Th17 and Th2) and CD25+ regulatory T cells (Treg), presented as percentage of live immune cells (PLICs) positive for CD45 ( n =3 analysed samples pooled from n =4–8 animals per sample). Flow cytometry analyses of ( g ) liver and ( h ) EWAT composition of total CD11c+ myeloid cells, F4/80+ myeloid cells, total macrophages (MΦ), CD11c+ MΦ and F4/80+ dendritic cells (DCs), presented as PLICs ( n =3 analysed samples pooled from n =4–8 animals per sample). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P
    Sil 6rα, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sil 6rα/product/R&D Systems
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sil 6rα - by Bioz Stars, 2021-05
    95/100 stars
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    93
    R&D Systems recombinant mouse il 6r
    Nerve injury upregulates <t>IL-6R</t> expression in adult mice DRG neurons. A , RT-qPCR shows a significant increase in mRNA expression of Il6ra gene in L4–L5 DRGs 3 d after sciatic nerve section ( n , number of experiments with duplicate measure of each gene; ** p
    Recombinant Mouse Il 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/recombinant mouse il 6r/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant mouse il 6r - by Bioz Stars, 2021-05
    93/100 stars
      Buy from Supplier

    94
    R&D Systems elisa
    Nerve injury upregulates <t>IL-6R</t> expression in adult mice DRG neurons. A , RT-qPCR shows a significant increase in mRNA expression of Il6ra gene in L4–L5 DRGs 3 d after sciatic nerve section ( n , number of experiments with duplicate measure of each gene; ** p
    Elisa, 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
    https://www.bioz.com/result/elisa/product/R&D Systems
    Average 94 stars, based on 1 article reviews
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    elisa - by Bioz Stars, 2021-05
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    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

    Journal: bioRxiv

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

    doi: 10.1101/2020.03.19.979104

    Figure Lengend 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

    Article Snippet: To restore IL-6 signaling of IL-6Rα KO DCs, human IL-6 (15ng/ml, 206-IL, R & D Systems) and soluble IL-6Rα (sIL-6Rα, 25ng/ml, 227-SR, R & D Systems) were added 1hr after LPS addition.

    Techniques: 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.

    Journal: bioRxiv

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

    doi: 10.1101/2020.03.19.979104

    Figure Lengend 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.

    Article Snippet: To restore IL-6 signaling of IL-6Rα KO DCs, human IL-6 (15ng/ml, 206-IL, R & D Systems) and soluble IL-6Rα (sIL-6Rα, 25ng/ml, 227-SR, R & D Systems) were added 1hr after LPS addition.

    Techniques: 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

    Journal: bioRxiv

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

    doi: 10.1101/2020.03.19.979104

    Figure Lengend 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

    Article Snippet: To restore IL-6 signaling of IL-6Rα KO DCs, human IL-6 (15ng/ml, 206-IL, R & D Systems) and soluble IL-6Rα (sIL-6Rα, 25ng/ml, 227-SR, R & D Systems) were added 1hr after LPS addition.

    Techniques: 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).

    Journal: bioRxiv

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

    doi: 10.1101/2020.03.19.979104

    Figure Lengend 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).

    Article Snippet: To restore IL-6 signaling of IL-6Rα KO DCs, human IL-6 (15ng/ml, 206-IL, R & D Systems) and soluble IL-6Rα (sIL-6Rα, 25ng/ml, 227-SR, R & D Systems) were added 1hr after LPS addition.

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

    Inflammatory profile of liver and EWAT (8-week HFD). ( a ) Liver ( n =10 versus 12) and ( b ) EWAT ( n =11) gene expression profile. Activity of c-Jun kinase (JNK) in ( c ) liver and ( d ) EWAT with the measurement of pS63 c-Jun by immunoblotting (representative immunoblots of 2 blots) and quantification ( n =8); relative unit (RU) to that of IL-6Rα f/f (Control). Flow cytometry analyses of ( e ) liver and ( f ) EWAT composition of total T cells, CD8+ and CD4+ T cells, T helper cells (Th1, Th17 and Th2) and CD25+ regulatory T cells (Treg), presented as percentage of live immune cells (PLICs) positive for CD45 ( n =3 analysed samples pooled from n =4–8 animals per sample). Flow cytometry analyses of ( g ) liver and ( h ) EWAT composition of total CD11c+ myeloid cells, F4/80+ myeloid cells, total macrophages (MΦ), CD11c+ MΦ and F4/80+ dendritic cells (DCs), presented as PLICs ( n =3 analysed samples pooled from n =4–8 animals per sample). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Inflammatory profile of liver and EWAT (8-week HFD). ( a ) Liver ( n =10 versus 12) and ( b ) EWAT ( n =11) gene expression profile. Activity of c-Jun kinase (JNK) in ( c ) liver and ( d ) EWAT with the measurement of pS63 c-Jun by immunoblotting (representative immunoblots of 2 blots) and quantification ( n =8); relative unit (RU) to that of IL-6Rα f/f (Control). Flow cytometry analyses of ( e ) liver and ( f ) EWAT composition of total T cells, CD8+ and CD4+ T cells, T helper cells (Th1, Th17 and Th2) and CD25+ regulatory T cells (Treg), presented as percentage of live immune cells (PLICs) positive for CD45 ( n =3 analysed samples pooled from n =4–8 animals per sample). Flow cytometry analyses of ( g ) liver and ( h ) EWAT composition of total CD11c+ myeloid cells, F4/80+ myeloid cells, total macrophages (MΦ), CD11c+ MΦ and F4/80+ dendritic cells (DCs), presented as PLICs ( n =3 analysed samples pooled from n =4–8 animals per sample). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Expressing, Activity Assay, Western Blot, Flow Cytometry, Cytometry, Two Tailed Test

    Augmented IL-6 signalling via sIL-6Rα after prolonged HFD feeding normalizes T cell functions in IL-6Rα T-KO (8- and 16-week HFD). ( a ) Compiled representative histograms and ( b ) quantification (MFI) of flow cytometry analyses of stimulated pY701 STAT-1 and pY705 STAT-3 with control (unstimulated), IL-6 (70 ng ml −1 ), IL-6—IL-6Rα complex (200 ng ml −1 ) and soluble IL-6Rα alone (130 ng ml −1 ) in enriched splenic T cell isolation from random-fed IL-6Rα f/f and IL-6Rα T-KO animals at 8 and 16 weeks of HFD feeding ( n =10–12 versus 10–12). ( c ) Flow cytometry analyses of T cell chemotaxis with control (no treatment), IL-6 (70 ng ml −1 ) and IL-6—IL-6Rα complex (200 ng ml −1 ), presented as percentage of cells pre-chemotaxis (PPC); enriched splenic T cells isolated from random-fed IL-6Rα f/f and IL-6Rα T-KO animals at 8 weeks ( n =6 versus 4) and 16 weeks ( n =4 versus 7) of HFD feeding. Two-way analysis of variance (ANOVA) with multiple analysis used for statistical analyses (* P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Augmented IL-6 signalling via sIL-6Rα after prolonged HFD feeding normalizes T cell functions in IL-6Rα T-KO (8- and 16-week HFD). ( a ) Compiled representative histograms and ( b ) quantification (MFI) of flow cytometry analyses of stimulated pY701 STAT-1 and pY705 STAT-3 with control (unstimulated), IL-6 (70 ng ml −1 ), IL-6—IL-6Rα complex (200 ng ml −1 ) and soluble IL-6Rα alone (130 ng ml −1 ) in enriched splenic T cell isolation from random-fed IL-6Rα f/f and IL-6Rα T-KO animals at 8 and 16 weeks of HFD feeding ( n =10–12 versus 10–12). ( c ) Flow cytometry analyses of T cell chemotaxis with control (no treatment), IL-6 (70 ng ml −1 ) and IL-6—IL-6Rα complex (200 ng ml −1 ), presented as percentage of cells pre-chemotaxis (PPC); enriched splenic T cells isolated from random-fed IL-6Rα f/f and IL-6Rα T-KO animals at 8 weeks ( n =6 versus 4) and 16 weeks ( n =4 versus 7) of HFD feeding. Two-way analysis of variance (ANOVA) with multiple analysis used for statistical analyses (* P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Flow Cytometry, Cytometry, Cell Isolation, Chemotaxis Assay, Isolation

    Improved glucose homeostasis and lipid metabolism in IL-6Rα T-KO (8-week HFD). ( a ) Blood glucose (BG) and glucose infusion rate (GIR) during hyperinsulinaemic-euglycaemic (HIEG) clamp experiments ( n =9); two-way analysis of variance (ANOVA) was performed on data from the green-shaded steady state (130–180 min). ( b ) Rate of hepatic glucose production (HGP) at basal and during steady state of HIEG clamp ( n =9). ( c ) Rate of glucose uptake (GU) during steady state of HIEG clamp ( n =9). Representative immunoblots of 3 blots and quantification of ( d ) liver and ( e ) EWAT pS473 Akt, total Akt and eEF2 at time 180 min during steady state of HIEG clamp ( n =9); relative unit (RU) to that of IL-6Rα f/f (Control). ( f ) Representative liver sections of 22 samples with haematoxylin and eosin (H E) staining; scale bars, 75 μm. ( g ) Hepatic content of triglycerides (TG), free cholesterols (Ch), cholesteryl esters (CE), diglycerides (DAG) and ceramides ( n =6). ( h ) Representative EWAT sections of 20 samples with F4/80 staining and quantification of adipocyte size in area and crown-like structure (CLS) counts ( n =8 versus 12); scale bars, 100 pixels. ( i ) Fast (12 h) and refeed (2 h) plasma TG and cholesterol levels ( n =19 versus 20). Two-tailed t -tests and two-way ANOVA used for statistical analyses (* P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Improved glucose homeostasis and lipid metabolism in IL-6Rα T-KO (8-week HFD). ( a ) Blood glucose (BG) and glucose infusion rate (GIR) during hyperinsulinaemic-euglycaemic (HIEG) clamp experiments ( n =9); two-way analysis of variance (ANOVA) was performed on data from the green-shaded steady state (130–180 min). ( b ) Rate of hepatic glucose production (HGP) at basal and during steady state of HIEG clamp ( n =9). ( c ) Rate of glucose uptake (GU) during steady state of HIEG clamp ( n =9). Representative immunoblots of 3 blots and quantification of ( d ) liver and ( e ) EWAT pS473 Akt, total Akt and eEF2 at time 180 min during steady state of HIEG clamp ( n =9); relative unit (RU) to that of IL-6Rα f/f (Control). ( f ) Representative liver sections of 22 samples with haematoxylin and eosin (H E) staining; scale bars, 75 μm. ( g ) Hepatic content of triglycerides (TG), free cholesterols (Ch), cholesteryl esters (CE), diglycerides (DAG) and ceramides ( n =6). ( h ) Representative EWAT sections of 20 samples with F4/80 staining and quantification of adipocyte size in area and crown-like structure (CLS) counts ( n =8 versus 12); scale bars, 100 pixels. ( i ) Fast (12 h) and refeed (2 h) plasma TG and cholesterol levels ( n =19 versus 20). Two-tailed t -tests and two-way ANOVA used for statistical analyses (* P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Western Blot, Staining, Two Tailed Test

    Physiological parameters and metabolic characterization of IL-6Rα T-KO mice (16-week HFD). ( a ) BW curve from 8 to 16 weeks on HFD and percentage of body weight gain ( n =22 versus 25). ( b ) Fasting (6 h) blood glucose ( n =22 versus 25), plasma insulin and HOMA-IR ( n =16 versus 19). ( c ) Blood glucose concentrations during IPGTT after a 6-h fast ( n =22 versus 25). ( d ) Blood glucose as a percentage of basal value during IPITT after a 2-h fast ( n =16 versus 21). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Physiological parameters and metabolic characterization of IL-6Rα T-KO mice (16-week HFD). ( a ) BW curve from 8 to 16 weeks on HFD and percentage of body weight gain ( n =22 versus 25). ( b ) Fasting (6 h) blood glucose ( n =22 versus 25), plasma insulin and HOMA-IR ( n =16 versus 19). ( c ) Blood glucose concentrations during IPGTT after a 6-h fast ( n =22 versus 25). ( d ) Blood glucose as a percentage of basal value during IPITT after a 2-h fast ( n =16 versus 21). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Mouse Assay, Two Tailed Test

    Inflammatory profile of liver and EWAT (16-week HFD). Flow cytometry analyses of ( a ) liver and ( b ) EWAT composition of total CD11c+ myeloid cells, F4/80+ myeloid cells, total MΦ, CD11c+ MΦ and F4/80+ DC, presented as PLICs ( n =3 analysed samples pooled from n =4–8 animals per sample). Flow cytometry analyses of ( c ) liver and ( d ) EWAT composition of total T cells, CD8+ and CD4+ T cells, Th1, Th17, Th2 and CD25+ Treg, presented as PLIC CD45+ ( n =3–5 analysed samples pooled from n =4–8 animals per sample); ND, not detected. JNK activity in ( e ) liver and ( f ) EWAT with the measurement of pS63 c-Jun by immunoblotting (representative immunoblots of 2 blots) and quantification ( n =8); relative unit (RU) to that of IL-6Rα f/f (Control). ( g ) Liver and ( h ) EWAT gene expression profile ( n =11). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Inflammatory profile of liver and EWAT (16-week HFD). Flow cytometry analyses of ( a ) liver and ( b ) EWAT composition of total CD11c+ myeloid cells, F4/80+ myeloid cells, total MΦ, CD11c+ MΦ and F4/80+ DC, presented as PLICs ( n =3 analysed samples pooled from n =4–8 animals per sample). Flow cytometry analyses of ( c ) liver and ( d ) EWAT composition of total T cells, CD8+ and CD4+ T cells, Th1, Th17, Th2 and CD25+ Treg, presented as PLIC CD45+ ( n =3–5 analysed samples pooled from n =4–8 animals per sample); ND, not detected. JNK activity in ( e ) liver and ( f ) EWAT with the measurement of pS63 c-Jun by immunoblotting (representative immunoblots of 2 blots) and quantification ( n =8); relative unit (RU) to that of IL-6Rα f/f (Control). ( g ) Liver and ( h ) EWAT gene expression profile ( n =11). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Flow Cytometry, Cytometry, Activity Assay, Western Blot, Expressing, Two Tailed Test

    Severe glucose homeostasis and lipid metabolism in IL-6Rα T-KO (16-week HFD). ( a ) BG and GIR during HIEG clamp experiments ( n =6 versus 9); two-way analysis of variance (ANOVA) was performed on data from the green-shaded steady state (130–180 min). ( b ) Rate of HGP at basal and during steady state of HIEG clamp ( n =6 versus 9). ( c ) Rate of GU during steady state of HIEG clamp ( n =6 versus 9). Representative immunoblots of 3 blots and quantification of ( d ) liver and ( e ) EWAT pS473 Akt, total Akt and eEF2 at time 180 min during steady state of HIEG clamp ( n =6 versus 9); relative unit (RU) to that of IL-6Rα f/f (Control). ( f ) Representative liver sections of 21 samples with haematoxylin and eosin (H E) staining; scale bars, 75 μm. ( g ) Hepatic content of TG, Ch, CE, DAG and ceramides ( n =9 versus 10). ( h ) Representative EWAT sections of 16 samples with F4/80 staining and quantification of adipocyte size in area and CLS counts ( n =7 versus 9); scale bars, 100 pixels. ( i ) Fast (12 h) and refeed (2 h) plasma TG and cholesterol levels ( n =16 versus 17). Two-tailed t -tests and two-way ANOVA used for statistical analyses (*** P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Severe glucose homeostasis and lipid metabolism in IL-6Rα T-KO (16-week HFD). ( a ) BG and GIR during HIEG clamp experiments ( n =6 versus 9); two-way analysis of variance (ANOVA) was performed on data from the green-shaded steady state (130–180 min). ( b ) Rate of HGP at basal and during steady state of HIEG clamp ( n =6 versus 9). ( c ) Rate of GU during steady state of HIEG clamp ( n =6 versus 9). Representative immunoblots of 3 blots and quantification of ( d ) liver and ( e ) EWAT pS473 Akt, total Akt and eEF2 at time 180 min during steady state of HIEG clamp ( n =6 versus 9); relative unit (RU) to that of IL-6Rα f/f (Control). ( f ) Representative liver sections of 21 samples with haematoxylin and eosin (H E) staining; scale bars, 75 μm. ( g ) Hepatic content of TG, Ch, CE, DAG and ceramides ( n =9 versus 10). ( h ) Representative EWAT sections of 16 samples with F4/80 staining and quantification of adipocyte size in area and CLS counts ( n =7 versus 9); scale bars, 100 pixels. ( i ) Fast (12 h) and refeed (2 h) plasma TG and cholesterol levels ( n =16 versus 17). Two-tailed t -tests and two-way ANOVA used for statistical analyses (*** P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Western Blot, Staining, Two Tailed Test

    Physiological parameters and metabolic characterization of IL-6Rα T-KO mice (8-week HFD). ( a ) Body weight (BW) curve on HFD and percentage of body weight gain ( n =21 versus 26). ( b ) Fasting (6 h) blood glucose ( n =21 versus 26), plasma insulin and HOMA-IR ( n =18 versus 16). ( c ) Blood glucose concentrations during intraperitoneal glucose tolerance tests (IPGTT) after a 6 h fast ( n =21 versus 26). ( d ) Blood glucose as a percentage of basal value during intraperitoneal insulin tolerance tests (IPITT) after a 2 h fast ( n =15 versus 18). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Physiological parameters and metabolic characterization of IL-6Rα T-KO mice (8-week HFD). ( a ) Body weight (BW) curve on HFD and percentage of body weight gain ( n =21 versus 26). ( b ) Fasting (6 h) blood glucose ( n =21 versus 26), plasma insulin and HOMA-IR ( n =18 versus 16). ( c ) Blood glucose concentrations during intraperitoneal glucose tolerance tests (IPGTT) after a 6 h fast ( n =21 versus 26). ( d ) Blood glucose as a percentage of basal value during intraperitoneal insulin tolerance tests (IPITT) after a 2 h fast ( n =15 versus 18). Two-tailed t -tests and two-way analysis of variance (ANOVA) used for statistical analyses (* P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Mouse Assay, Two Tailed Test

    Increased IL-6 and sIL-6Rα levels upon prolonged HFD feeding (8- and 16-week HFD). ( a ) Serum IL-6 levels ( n =11 versus 10–14). ( b ) Serum sIL-6Rα levels ( n =20 versus 20). ( c ) Liver IL-6 content ( n =11–12 versus 11–12). ( d ) EWAT IL-6 content ( n =11–12 versus 11–12). Representative immunoblots and quantification of ( e ) liver ( n =8 versus 8) and ( f ) EWAT sIL-6Rα and eEF2 ( n =4–8 versus 4–8) with a serum sample (S); relative unit (RU) to that of IL-6Rα f/f (Control). Flow cytometry analyses of gp130 expression in cells isolated from ( g ) liver ( n =10–12 versus 10–12) and ( h ) EWAT ( n =10–12 versus 10–13), presented as gp130+ cells in percentage of live CD3+ immune cells (PLIC CD3+) and as amount of gp130 detection in median fluorescence intensity per positive cell (MFI/+Cell). Two-way analysis of variance (ANOVA) with multiple analysis used for statistical analyses (* P

    Journal: Nature Communications

    Article Title: Temporal and tissue-specific requirements for T-lymphocyte IL-6 signalling in obesity-associated inflammation and insulin resistance

    doi: 10.1038/ncomms14803

    Figure Lengend Snippet: Increased IL-6 and sIL-6Rα levels upon prolonged HFD feeding (8- and 16-week HFD). ( a ) Serum IL-6 levels ( n =11 versus 10–14). ( b ) Serum sIL-6Rα levels ( n =20 versus 20). ( c ) Liver IL-6 content ( n =11–12 versus 11–12). ( d ) EWAT IL-6 content ( n =11–12 versus 11–12). Representative immunoblots and quantification of ( e ) liver ( n =8 versus 8) and ( f ) EWAT sIL-6Rα and eEF2 ( n =4–8 versus 4–8) with a serum sample (S); relative unit (RU) to that of IL-6Rα f/f (Control). Flow cytometry analyses of gp130 expression in cells isolated from ( g ) liver ( n =10–12 versus 10–12) and ( h ) EWAT ( n =10–12 versus 10–13), presented as gp130+ cells in percentage of live CD3+ immune cells (PLIC CD3+) and as amount of gp130 detection in median fluorescence intensity per positive cell (MFI/+Cell). Two-way analysis of variance (ANOVA) with multiple analysis used for statistical analyses (* P

    Article Snippet: Detection of STAT-1/3 phosphorylation by flow cytometry Following the manufacturer's guidelines of BD Phosflow Protocol II, the mild alcohol method (BD Biosciences), 106 enriched splenic T cells in 100 μl per well on a conical-bottomed 96-well plate were cultured for 30 min in serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) before being transferred to another plate with control SF medium (unstimulated), SF media containing freshly prepared 70 ng ml−1 IL-6 (406-ML-005, R & D Systems), 200 ng ml−1 IL-6—IL-6Rα complex (9038-SR-025, R & D Systems) or 130 ng ml−1 sIL-6Rα alone (1830-SR-025, R & D Systems).

    Techniques: Western Blot, Flow Cytometry, Cytometry, Expressing, Isolation, Fluorescence

    Nerve injury upregulates IL-6R expression in adult mice DRG neurons. A , RT-qPCR shows a significant increase in mRNA expression of Il6ra gene in L4–L5 DRGs 3 d after sciatic nerve section ( n , number of experiments with duplicate measure of each gene; ** p

    Journal: The Journal of Neuroscience

    Article Title: An Autocrine Neuronal Interleukin-6 Loop Mediates Chloride Accumulation and NKCC1 Phosphorylation in Axotomized Sensory Neurons

    doi: 10.1523/JNEUROSCI.3382-11.2011

    Figure Lengend Snippet: Nerve injury upregulates IL-6R expression in adult mice DRG neurons. A , RT-qPCR shows a significant increase in mRNA expression of Il6ra gene in L4–L5 DRGs 3 d after sciatic nerve section ( n , number of experiments with duplicate measure of each gene; ** p

    Article Snippet: Blocking-function mouse IL-6 antibody and recombinant mouse IL-6R [soluble IL-6R obtained from a DNA sequence encoding the extracellular domain of mouse IL-6R ( )] were from R & D Systems.

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR

    IL-6R is expressed in a subset of myelinated sensory neurons. A , Apotome images of transversal slices of axotomized DRG double stained with anti-IL-6R (red) and anti-NF-200 (green) antibodies. The merge image shows colocalization of IL-6R with NF-200-positive sensory neurons. B , Double immunochemistry using anti-TrkB antibody (green) shows IL-6R colocalization with TrkB-positive neurons (merge). C , Double immunochemistry using anti-Parvalbumin antibody (green) shows IL-6R colocalization with parvalbumin-positive neurons (merge). D , Double immunochemistry using anti-TrkA antibody (green) demonstrates no IL6-R colocalization with TrkA-positive neurons (merge). Scale bar, 30 μm.

    Journal: The Journal of Neuroscience

    Article Title: An Autocrine Neuronal Interleukin-6 Loop Mediates Chloride Accumulation and NKCC1 Phosphorylation in Axotomized Sensory Neurons

    doi: 10.1523/JNEUROSCI.3382-11.2011

    Figure Lengend Snippet: IL-6R is expressed in a subset of myelinated sensory neurons. A , Apotome images of transversal slices of axotomized DRG double stained with anti-IL-6R (red) and anti-NF-200 (green) antibodies. The merge image shows colocalization of IL-6R with NF-200-positive sensory neurons. B , Double immunochemistry using anti-TrkB antibody (green) shows IL-6R colocalization with TrkB-positive neurons (merge). C , Double immunochemistry using anti-Parvalbumin antibody (green) shows IL-6R colocalization with parvalbumin-positive neurons (merge). D , Double immunochemistry using anti-TrkA antibody (green) demonstrates no IL6-R colocalization with TrkA-positive neurons (merge). Scale bar, 30 μm.

    Article Snippet: Blocking-function mouse IL-6 antibody and recombinant mouse IL-6R [soluble IL-6R obtained from a DNA sequence encoding the extracellular domain of mouse IL-6R ( )] were from R & D Systems.

    Techniques: Staining

    Il-6 regulates the expression of phospho-NKCC1 close to the plasma membrane of axotomized DRG neurons. A , Transverse section of L4–L6 DRG immunolabeled with an antibody generated against phospho-NKCC1; no signal is observed. B , Five days after nerve injury, phospho-NKCC1 staining is present at/over the plasma membrane in a subset of sensory neurons (arrows). C , Similar to WT DRG, phospho-NKCC1 staining was absent in transverse sections of L4–L6 DRGs from IL6 −/− mice. D , Five days after nerve injury, only a few neurons have a plasma membrane showing phospho-NKCC1 staining in DRG from IL-6 −/− mice (arrow). E , Confocal microscopy of double-immunofluorescent labeling for phospho-NKCC1 (p-NKCC1) and glutamine synthase (GS) shows no coexpression. F , Confocal microscopy of double-immunofluorescent labeling for p-NKCC1 and IL-6R shows expression in the same cell type. Merge image shows a co expression that appears as an alternate punctiform staining at the plasma level. Scale bars, 30 μm. Insets in merge images are 3× magnifications of the region marked by white asterisk. A–D , 20× objective; E , F , 40× objective.

    Journal: The Journal of Neuroscience

    Article Title: An Autocrine Neuronal Interleukin-6 Loop Mediates Chloride Accumulation and NKCC1 Phosphorylation in Axotomized Sensory Neurons

    doi: 10.1523/JNEUROSCI.3382-11.2011

    Figure Lengend Snippet: Il-6 regulates the expression of phospho-NKCC1 close to the plasma membrane of axotomized DRG neurons. A , Transverse section of L4–L6 DRG immunolabeled with an antibody generated against phospho-NKCC1; no signal is observed. B , Five days after nerve injury, phospho-NKCC1 staining is present at/over the plasma membrane in a subset of sensory neurons (arrows). C , Similar to WT DRG, phospho-NKCC1 staining was absent in transverse sections of L4–L6 DRGs from IL6 −/− mice. D , Five days after nerve injury, only a few neurons have a plasma membrane showing phospho-NKCC1 staining in DRG from IL-6 −/− mice (arrow). E , Confocal microscopy of double-immunofluorescent labeling for phospho-NKCC1 (p-NKCC1) and glutamine synthase (GS) shows no coexpression. F , Confocal microscopy of double-immunofluorescent labeling for p-NKCC1 and IL-6R shows expression in the same cell type. Merge image shows a co expression that appears as an alternate punctiform staining at the plasma level. Scale bars, 30 μm. Insets in merge images are 3× magnifications of the region marked by white asterisk. A–D , 20× objective; E , F , 40× objective.

    Article Snippet: Blocking-function mouse IL-6 antibody and recombinant mouse IL-6R [soluble IL-6R obtained from a DNA sequence encoding the extracellular domain of mouse IL-6R ( )] were from R & D Systems.

    Techniques: Expressing, Immunolabeling, Generated, Staining, Mouse Assay, Confocal Microscopy, Labeling