rmil 6  (PeproTech)

 
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    Name:
    Recombinant Murine IL 6
    Description:
    IL 6 is a pleiotropic cytokine that plays an important role in host defense by regulating immune and inflammatory responses Produced by T cells monocytes fibroblasts endothelial cells and keratinocytes IL 6 has diverse biological functions It stimulates B cell differentiation and antibody production synergizes with IL 3 in megakaryocyte development and platelet production induces expression of hepatic acute phase proteins and regulates bone metabolism IL 6 signals through the IL 6 receptor system that consists of two chains IL 6R α and gp130 Murine IL 6 is inactive on human cells while both human and murine are equally active on murine cells Recombinant murine IL 6 is a 21 7 kDa protein containing 188 amino acid residues
    Catalog Number:
    216-16-100UG
    Price:
    860.00
    Category:
    Recombinant Proteins
    Source:
    E.coli
    Reactivity:
    Monkey
    Purity:
    98.0
    Quantity:
    100UG
    Buy from Supplier


    Structured Review

    PeproTech rmil 6
    Effect of alpinetin on the imbalance of Th17/Treg. a – d Mice were orally administrated of 2.5% DSS for 7 days, and followed by sterile distilled water alone for another 3 days. The alpinetin (7.5, 15, 30 mg/kg) and 5-aminosalicylic acid (5-ASA, 200 mg/kg) were orally administered daily for consecutive 10 days. Then, mice were sacrificed, and colons were collected. The protein levels of IL-17 and IL-10 in colons were detected by using ELISA ( a ); the mRNA levels of IL-17, IL-10, RORγt and Foxp3 were detected by using Q-PCR assay ( b , c ); the percentages of Th17 cells and Treg cells in mesenteric lymph nodes (MLNs) and colonic lamina proprias (LPs) were detected by using flow cytometry assay ( d , e ). f CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL), rhTGF-β1 (5 ng/mL), <t>rmIL-6</t> (20 ng/mL), rmIL-2 (300 IU/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Th17 cells were detected by using flow cytometry assay. In addition, CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Treg cells were detected by using flow cytometry assay. g CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, mRNA and protein levels of IL-10 were detected by using Q-PCR and ELISA, respectively. h CD4 + T cells were cultured with alpinetin (1, 3, 10, 30, 100 μM) for 72 h, the viability and proliferation of CD4 + T cells were detected by using MTT and CCK8 assays, respectively. The data were presented as means ± S.E.M. of three independent experiments or six mice in each group. ## P
    IL 6 is a pleiotropic cytokine that plays an important role in host defense by regulating immune and inflammatory responses Produced by T cells monocytes fibroblasts endothelial cells and keratinocytes IL 6 has diverse biological functions It stimulates B cell differentiation and antibody production synergizes with IL 3 in megakaryocyte development and platelet production induces expression of hepatic acute phase proteins and regulates bone metabolism IL 6 signals through the IL 6 receptor system that consists of two chains IL 6R α and gp130 Murine IL 6 is inactive on human cells while both human and murine are equally active on murine cells Recombinant murine IL 6 is a 21 7 kDa protein containing 188 amino acid residues
    https://www.bioz.com/result/rmil 6/product/PeproTech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rmil 6 - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation"

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0814-4

    Effect of alpinetin on the imbalance of Th17/Treg. a – d Mice were orally administrated of 2.5% DSS for 7 days, and followed by sterile distilled water alone for another 3 days. The alpinetin (7.5, 15, 30 mg/kg) and 5-aminosalicylic acid (5-ASA, 200 mg/kg) were orally administered daily for consecutive 10 days. Then, mice were sacrificed, and colons were collected. The protein levels of IL-17 and IL-10 in colons were detected by using ELISA ( a ); the mRNA levels of IL-17, IL-10, RORγt and Foxp3 were detected by using Q-PCR assay ( b , c ); the percentages of Th17 cells and Treg cells in mesenteric lymph nodes (MLNs) and colonic lamina proprias (LPs) were detected by using flow cytometry assay ( d , e ). f CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL), rhTGF-β1 (5 ng/mL), rmIL-6 (20 ng/mL), rmIL-2 (300 IU/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Th17 cells were detected by using flow cytometry assay. In addition, CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Treg cells were detected by using flow cytometry assay. g CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, mRNA and protein levels of IL-10 were detected by using Q-PCR and ELISA, respectively. h CD4 + T cells were cultured with alpinetin (1, 3, 10, 30, 100 μM) for 72 h, the viability and proliferation of CD4 + T cells were detected by using MTT and CCK8 assays, respectively. The data were presented as means ± S.E.M. of three independent experiments or six mice in each group. ## P
    Figure Legend Snippet: Effect of alpinetin on the imbalance of Th17/Treg. a – d Mice were orally administrated of 2.5% DSS for 7 days, and followed by sterile distilled water alone for another 3 days. The alpinetin (7.5, 15, 30 mg/kg) and 5-aminosalicylic acid (5-ASA, 200 mg/kg) were orally administered daily for consecutive 10 days. Then, mice were sacrificed, and colons were collected. The protein levels of IL-17 and IL-10 in colons were detected by using ELISA ( a ); the mRNA levels of IL-17, IL-10, RORγt and Foxp3 were detected by using Q-PCR assay ( b , c ); the percentages of Th17 cells and Treg cells in mesenteric lymph nodes (MLNs) and colonic lamina proprias (LPs) were detected by using flow cytometry assay ( d , e ). f CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL), rhTGF-β1 (5 ng/mL), rmIL-6 (20 ng/mL), rmIL-2 (300 IU/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Th17 cells were detected by using flow cytometry assay. In addition, CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Treg cells were detected by using flow cytometry assay. g CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, mRNA and protein levels of IL-10 were detected by using Q-PCR and ELISA, respectively. h CD4 + T cells were cultured with alpinetin (1, 3, 10, 30, 100 μM) for 72 h, the viability and proliferation of CD4 + T cells were detected by using MTT and CCK8 assays, respectively. The data were presented as means ± S.E.M. of three independent experiments or six mice in each group. ## P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Flow Cytometry, Cytometry, Cell Culture, MTT Assay

    2) Product Images from "Cytokine Enhancement of DNA Immunization Leads to Effective Treatment of Established Pulmonary Metastases"

    Article Title: Cytokine Enhancement of DNA Immunization Leads to Effective Treatment of Established Pulmonary Metastases

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

    doi:

    Active immunotherapy of established pulmonary metastases with the pCMV/ β -gal vaccine plus systemic administration of rhIL-2, rmIL-6, or rhIL-7. BALB/c mice were injected i.v. with 5 × 10 5 CT26.CL25 ( β gal + ) tumor cells. On day 2 following tumor challenge, treated mice were immunized with 10 μ g of pCMV/ β -gal. Each mouse received 10 shots of 0.5 mg of gold, each shot delivering 1 μ g of DNA. On day 3, mice (5 to 10 mice per group) began regimens of i.p. cytokine injections as described in Materials and Methods . On day 12, lungs were harvested and the pulmonary metastases were enumerated in a coded, blind fashion. Nonparametric statistical analysis was done using the Kruskal-Wallis test. The group with asterisks above were found to be statistically significant compared with either cytokine alone (*, p 2 = 0.001; **, p 2 = 0.003; ***, p 2 = 0.0002) or to β -gal DNA alone (*, p 2 = 0.003; **, p 2 = 0.0003; ***, p 2 = 0.001). The graph represents a summary of two experiments.
    Figure Legend Snippet: Active immunotherapy of established pulmonary metastases with the pCMV/ β -gal vaccine plus systemic administration of rhIL-2, rmIL-6, or rhIL-7. BALB/c mice were injected i.v. with 5 × 10 5 CT26.CL25 ( β gal + ) tumor cells. On day 2 following tumor challenge, treated mice were immunized with 10 μ g of pCMV/ β -gal. Each mouse received 10 shots of 0.5 mg of gold, each shot delivering 1 μ g of DNA. On day 3, mice (5 to 10 mice per group) began regimens of i.p. cytokine injections as described in Materials and Methods . On day 12, lungs were harvested and the pulmonary metastases were enumerated in a coded, blind fashion. Nonparametric statistical analysis was done using the Kruskal-Wallis test. The group with asterisks above were found to be statistically significant compared with either cytokine alone (*, p 2 = 0.001; **, p 2 = 0.003; ***, p 2 = 0.0002) or to β -gal DNA alone (*, p 2 = 0.003; **, p 2 = 0.0003; ***, p 2 = 0.001). The graph represents a summary of two experiments.

    Techniques Used: Mouse Assay, Injection

    3) Product Images from "Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation"

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0814-4

    Effect of alpinetin on the imbalance of Th17/Treg. a – d Mice were orally administrated of 2.5% DSS for 7 days, and followed by sterile distilled water alone for another 3 days. The alpinetin (7.5, 15, 30 mg/kg) and 5-aminosalicylic acid (5-ASA, 200 mg/kg) were orally administered daily for consecutive 10 days. Then, mice were sacrificed, and colons were collected. The protein levels of IL-17 and IL-10 in colons were detected by using ELISA ( a ); the mRNA levels of IL-17, IL-10, RORγt and Foxp3 were detected by using Q-PCR assay ( b , c ); the percentages of Th17 cells and Treg cells in mesenteric lymph nodes (MLNs) and colonic lamina proprias (LPs) were detected by using flow cytometry assay ( d , e ). f CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL), rhTGF-β1 (5 ng/mL), rmIL-6 (20 ng/mL), rmIL-2 (300 IU/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Th17 cells were detected by using flow cytometry assay. In addition, CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Treg cells were detected by using flow cytometry assay. g CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, mRNA and protein levels of IL-10 were detected by using Q-PCR and ELISA, respectively. h CD4 + T cells were cultured with alpinetin (1, 3, 10, 30, 100 μM) for 72 h, the viability and proliferation of CD4 + T cells were detected by using MTT and CCK8 assays, respectively. The data were presented as means ± S.E.M. of three independent experiments or six mice in each group. ## P
    Figure Legend Snippet: Effect of alpinetin on the imbalance of Th17/Treg. a – d Mice were orally administrated of 2.5% DSS for 7 days, and followed by sterile distilled water alone for another 3 days. The alpinetin (7.5, 15, 30 mg/kg) and 5-aminosalicylic acid (5-ASA, 200 mg/kg) were orally administered daily for consecutive 10 days. Then, mice were sacrificed, and colons were collected. The protein levels of IL-17 and IL-10 in colons were detected by using ELISA ( a ); the mRNA levels of IL-17, IL-10, RORγt and Foxp3 were detected by using Q-PCR assay ( b , c ); the percentages of Th17 cells and Treg cells in mesenteric lymph nodes (MLNs) and colonic lamina proprias (LPs) were detected by using flow cytometry assay ( d , e ). f CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL), rhTGF-β1 (5 ng/mL), rmIL-6 (20 ng/mL), rmIL-2 (300 IU/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Th17 cells were detected by using flow cytometry assay. In addition, CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, percentages of Treg cells were detected by using flow cytometry assay. g CD4 + T cells were treated with anti-CD3/CD28 (2 μg/mL) and alpinetin (1, 3, 10, 30 μM) for 72 h, mRNA and protein levels of IL-10 were detected by using Q-PCR and ELISA, respectively. h CD4 + T cells were cultured with alpinetin (1, 3, 10, 30, 100 μM) for 72 h, the viability and proliferation of CD4 + T cells were detected by using MTT and CCK8 assays, respectively. The data were presented as means ± S.E.M. of three independent experiments or six mice in each group. ## P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Flow Cytometry, Cytometry, Cell Culture, MTT Assay

    Related Articles

    Recombinant:

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    Article Snippet: 2.5×105 T cells and 7.5×105 APCs were plated into 96-well round bottom plates and stimulated with the P14 GP33–41 peptide (300 ng/ml) in Iscove’s modified Dulbecco’s medium (cIMDM) supplemented with 10% FBS (vol/vol), penicillin, streptomycin, gentamycin, and 50 nM 2-mercaptoethanol. .. Culture conditions, as indicated, included 10 ng/ml recombinant IL-23 (eBioscience), 20 ng/ml recombinant IL-6 (PeproTech, Inc.), 10 ng/ml recombinant IL-12/23p40 homodimer, 5 ng/ml rhTGF-β1, 5 ng/ml of recombinant IL-12, all from R & D Systems, 10 µg/ml anti-mouse/human TGF-β1 (clone 1D11.16.8) and 10 µg/ml anti-mouse IL-6R (clone 15A7), both from Bio-X-Cell. ..

    Article Title: Vagus-macrophage-hepatocyte link promotes post-injury liver regeneration and whole-body survival through hepatic FoxM1 activation
    Article Snippet: .. At 48 h after isolation, hepatocytes were incubated with 100 ng/ml recombinant murine IL-6 (PEPROTECH, London, UK) for 6 h, and then collected for RNA extraction. .. In experiments employing ex vivo carbachol treatment, to avoid macrophage contamination, hepatocytes were incubated with 1 mM clodronate liposomes for 1 h starting at 24 h after isolation.

    Article Title: TLR4-induced NF-κB and MAPK signaling regulate the IL-6 mRNA stabilizing protein Arid5a
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    Enzyme-linked Immunosorbent Assay:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Purification:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Isolation:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Article Title: Vagus-macrophage-hepatocyte link promotes post-injury liver regeneration and whole-body survival through hepatic FoxM1 activation
    Article Snippet: .. At 48 h after isolation, hepatocytes were incubated with 100 ng/ml recombinant murine IL-6 (PEPROTECH, London, UK) for 6 h, and then collected for RNA extraction. .. In experiments employing ex vivo carbachol treatment, to avoid macrophage contamination, hepatocytes were incubated with 1 mM clodronate liposomes for 1 h starting at 24 h after isolation.

    MTT Assay:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Quantitative RT-PCR:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Activity Assay:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    SYBR Green Assay:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Methylation:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Chromatin Immunoprecipitation:

    Article Title: Alpinetin exerts anti-colitis efficacy by activating AhR, regulating miR-302/DNMT-1/CREB signals, and therefore promoting Treg differentiation
    Article Snippet: Chemicals and reagents Alpinetin (C16 H14 O4 , MW: 270.28, purity > 98%) was purchased from Jingzhu biological technology (Nanjing, China) ; 5-ASA sustained release granules were purchased from Ipsen Pharma (Houdan, France); TCDD was purchased from J & K Chemical (Beijing, China); CH223191 was purchased from Selleckchem (Houston, USA); fetal bovine serum (FBS) was purchased from PAA (Linz, Germany); DSS (36–50 kDa) was purchased from MP Biomedical (Aurora, USA); MPO activity detection kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); nuclear and cytoplasmic protein extraction kit were purchased from Nanjing KeyGen Biotech. .. Inc. (Nanjing, China); IL-1β, TNF-α, IL-10 and IL-17A enzyme-linked immune sorbent assay (ELISA) kits were purchased from Dakewe Biotech (Shenzhen, China); FITC-anti-CD4, APC-anti-CD25, PE-anti-Foxp3, PE-anti-IL-17A, IgG2a K isotype control PE, fixation/permeabilization concentrate, diluent reagent, purified anti-mouse CD3e and anti-mouse CD28 mAbs were purchased from eBioscience (San Diego, USA); PMA/Ionomycin mixture and BFA/Monensin mixture were purchased from MultiSciences Biotech (Hangzhou, China); Mouse CD4+ CD62L+ T-cell isolation kit II was purchased from Miltenyi Biotech (Cologne, Germany); rmIL-2, rmIL-6 and rhTGF-β1 were purchased from PeproTech (Madison, USA); MTT, lipofectamine TM 2000 and TRIzol reagent were purchased from Invitrogen (Carlsbad, USA); anti-CYP1A1 antibody, anti-β-actin antibody, anti-lamin B antibody, protein A + G agarose beads, peroxidase-conjugated secondary antibody and pcDNA3.1(+)-mDNMT-1 were purchased from Bioworld (Georgia, USA); siAhR was purchased from Genechem (Shanghai, China); the Bulgen-loopTM miRNA qRT-PCR Primer Sets specific for miR-31, miR-219, miR-302, miR-148a, miR-21 and miR-155 were purchased from RiboBio (Guangzhou, China); EROD activity detection kit was purchased from Shanghai genmed Pharmaceutical Technology Co. Ltd. (Shanghai, China); HiScript™QRTSuperMix and AceQ™qPCR SYBR® Green Master Mix were purchased from Vazyme Biotech (Piscataway, USA); EZ DNA Methylation-Gold™ Kit was purchased from Zymo Research Corporation (CA, USA); genomic DNA Mini Preparation Kit, ChIP assay Kit, proteinase K and NP40 buffer were purchased from Beyotime Biotech (Nanjing, China); anti-DNMT-1 antibody was purchased from Abcam (Cambridge, UK); anti-CREB antibody was purchased from Cell Signaling Technology (MA, USA); anti-Foxp3 antibody, anti-AhR antibody, anti-HSP90 antibody and anti-ARNT antibody were purchased from Santa Cruz Biotechnology (CA, USA); ECL reagent was purchased from DiZhao Biotech (Shanghai, China). .. In vivo, TCDD and CH223191 were diluted in corn oil; alpinetin and 5-ASA were diluted in 0.5% carboxymethyl cellulose sodium-Na (CMC-Na).

    Incubation:

    Article Title: Vagus-macrophage-hepatocyte link promotes post-injury liver regeneration and whole-body survival through hepatic FoxM1 activation
    Article Snippet: .. At 48 h after isolation, hepatocytes were incubated with 100 ng/ml recombinant murine IL-6 (PEPROTECH, London, UK) for 6 h, and then collected for RNA extraction. .. In experiments employing ex vivo carbachol treatment, to avoid macrophage contamination, hepatocytes were incubated with 1 mM clodronate liposomes for 1 h starting at 24 h after isolation.

    RNA Extraction:

    Article Title: Vagus-macrophage-hepatocyte link promotes post-injury liver regeneration and whole-body survival through hepatic FoxM1 activation
    Article Snippet: .. At 48 h after isolation, hepatocytes were incubated with 100 ng/ml recombinant murine IL-6 (PEPROTECH, London, UK) for 6 h, and then collected for RNA extraction. .. In experiments employing ex vivo carbachol treatment, to avoid macrophage contamination, hepatocytes were incubated with 1 mM clodronate liposomes for 1 h starting at 24 h after isolation.

    Injection:

    Article Title: Circulating IL-6 mediates lung injury via CXCL1 production after acute kidney injury in mice
    Article Snippet: IL-6-deficient mice (Jackson Labs Strain B6.129S2-IL6/J) underwent ischemic AKI surgery as described above. .. Two hundred nanograms of recombinant murine IL-6 (PeproTech #216–16) in 200 μl of PBS with 0.1% BSA were injected at 1, 2, and 3 h after AKI for a total dose of 600 ng. .. Vehicle-treated mice received the same volume of PBS in 0.1% bovine serum albumin (BSA) at the same time points.

    Modification:

    Article Title: Sp3 is involved in the regulation of SOCS3 gene expression
    Article Snippet: The results indicate that this GC-rich element is involved in basal and induced transcriptional activation of the SOCS3 promoter and that Sp3 (specificity protein 3), a ubiquitously expressed transcription factor, but not Sp1 binds to this GC-rich motif. .. High prime labelling kit, Taq polymerase and recombinant Epo (erythropoietin) were purchased from Roche Molecular Biochemicals (Mannheim, Germany); restriction enzymes were obtained from New England Biolabs (Frankfurt, Germany); oligonucleotides were obtained from MWG-Biotech (Ebersberg, Germany); DMEM (Dulbecco's modified Eagle's medium), DMEM nutritional mix F12 and Opti-MEM were from Invitrogen (Karlsruhe, Germany); recombinant human IL-6 was a gift from Dr P. C. Heinrich (Institute of Biochemistry, University of Aachen, Aachen, Germany); murine IL-6 was from PeproTech; and fetal calf serum was from Perbio Science. .. The One-Step RT (reverse transcriptase)–PCR kit was from Qiagen (Hilden, Germany).

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  • 96
    PeproTech recombinant mouse il 6
    Immunologic alterations in tumor-bearing mice treated with chemotherapy and/or anti-BTLA Ab. a The percentages of CD223 expression of CD4 + T lymphocytes in splenocytes of various therapeutic groups. The percentage of CD223 + CD4 + T lymphocytes in splenocytes was highest in paclitaxel combined with anti-BTLA Ab 20 μg/mouse group ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) b The percentages of CD223 expression of CD8 + T lymphocytes in splenocytes of various therapeutic groups. The percentage of CD223 + CD8 + T lymphocytes in splenocytes was also highest with paclitaxel combined with anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) c The percentages of CD223 expression of CD4 + T lymphocytes in TACs of ascites of various therapeutic groups. The percentage of CD223 + CD4 + T lymphocytes in TACs of ascites was highest with paclitaxel and anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) d The percentages of CD223 expression of CD8 + T lymphocytes in TACs of ascites of various therapeutic groups. The percentage of CD223 + CD8 + T lymphocytes in TACs of ascites was also highest with paclitaxel combined with anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) e Tumoricidal activity of splenocytes of tumor-bearing mice receiving chemotherapy treated with/without anti-BTLA Ab in vitro. e1 Representative luminescence figures of the in vitro tumor killing abilities of splenocytes by the IVIS system. (5 mice in each group) e2 Quantification of luminescence of in vitro tumor killing abilities of splenocytes by the IVIS system. Compared with the luminescence of WF-3/Luc cells co-cultured with splenocytes without anti-BTLA Ab, less luminal activity was detected in WF-3/Luc cells co-cultured with splenocytes receiving in vitro anti-BTLA Ab ( p = 0.021 for WF-3/Luc:splenocyte = 1:100; p = 0.027 for WF-3/Luc:splenocyte = 1:50; and p = 0.039 for WF-3/Luc:splenocyte = 1:10, Kruskal-Wallis test). The splenocytes treated with anti-BTLA Ab could generate higher tumor killing activities than those without anti-BTLA Ab. (5 mice in each group) f Bar figures of concentrations of various cytokines in ascites of various groups. Note : F1: IL-12; F2: TNF-α; F3: IFN-γ; F4: <t>IL-6;</t> F5: IL-10; and F6: TGF-β. The pro-inflammatory cytokines such as IL-12 ( p = 0.002), TNF-α ( p = 0.002), and IFN-γ ( p = 0.001) were highest with the chemotherapy combined with anti-BTLA Ab 20 μg/mouse. The concentrations of ant-inflammatory cytokines such as IL-6 ( p = 0.83), IL-10 ( p = 0.85), and TGF-β ( p = 0.84) did not show differences among the various groups (all statistical analyses by Kruskal-Wallis test). (5 mice in each group)
    Recombinant Mouse Il 6, supplied by PeproTech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant mouse il 6/product/PeproTech
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant mouse il 6 - by Bioz Stars, 2021-05
    96/100 stars
      Buy from Supplier

    99
    PeproTech recombinant murine il 6
    MKP-1 signaling regulates Arid5a mRNA stability by inducing the translocalization of AUF-1 from the nucleus to the cytoplasm. ( A and B ) Analysis of MKP-1 mRNA and protein levels in MEF cells that were transfected with control siRNA and MKP-1 siRNA. ( C ) Quantitative real-time PCR analysis of AUF-1 mRNA levels in control and MKP-1 knockdown cells that were stimulated with LPS (3 μg/ml). ( D ) AUF-1 (p45, p42, p40, p37) levels in the cytoplasm versus the nucleus in control and MKP-1 knockdown MEF cells that were stimulated using LPS for 0–4 h. β-tubulin and Lamin A/C were used as the loading controls for cytoplasmic proteins and nuclear proteins, respectively. ( E ) Quantitative real-time PCR analysis of mRNA levels of Arid5a in control and MKP-1 knockdown MEFs that were stimulated with LPS and then treated with Actinomycin D for 0–90 min. ( F and G ) Quantitative real-time PCR analysis of mRNA levels of Arid5a and <t>IL-6</t> in control and MKP-1 knockdown MEFs that were stimulated with LPS. ( H ) Schematic diagram illustrating the post-transcriptional regulation of Arid5a mRNA. LPS signaling induces MKP-1, which in turn signals AUF-1 to translocate from the nucleus to the cytoplasm. In the cytoplasm, AUF-1 binds to AU-rich elements on the Arid5a 3΄ UTR to destabilize the Arid5a mRNA. This results in the inhibition of the stabilization of the IL-6 mRNA. All data are shown as the mean ± SD of three independent experiments. N.S., not significant. * P
    Recombinant Murine Il 6, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant murine il 6/product/PeproTech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant murine il 6 - by Bioz Stars, 2021-05
    99/100 stars
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    Immunologic alterations in tumor-bearing mice treated with chemotherapy and/or anti-BTLA Ab. a The percentages of CD223 expression of CD4 + T lymphocytes in splenocytes of various therapeutic groups. The percentage of CD223 + CD4 + T lymphocytes in splenocytes was highest in paclitaxel combined with anti-BTLA Ab 20 μg/mouse group ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) b The percentages of CD223 expression of CD8 + T lymphocytes in splenocytes of various therapeutic groups. The percentage of CD223 + CD8 + T lymphocytes in splenocytes was also highest with paclitaxel combined with anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) c The percentages of CD223 expression of CD4 + T lymphocytes in TACs of ascites of various therapeutic groups. The percentage of CD223 + CD4 + T lymphocytes in TACs of ascites was highest with paclitaxel and anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) d The percentages of CD223 expression of CD8 + T lymphocytes in TACs of ascites of various therapeutic groups. The percentage of CD223 + CD8 + T lymphocytes in TACs of ascites was also highest with paclitaxel combined with anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) e Tumoricidal activity of splenocytes of tumor-bearing mice receiving chemotherapy treated with/without anti-BTLA Ab in vitro. e1 Representative luminescence figures of the in vitro tumor killing abilities of splenocytes by the IVIS system. (5 mice in each group) e2 Quantification of luminescence of in vitro tumor killing abilities of splenocytes by the IVIS system. Compared with the luminescence of WF-3/Luc cells co-cultured with splenocytes without anti-BTLA Ab, less luminal activity was detected in WF-3/Luc cells co-cultured with splenocytes receiving in vitro anti-BTLA Ab ( p = 0.021 for WF-3/Luc:splenocyte = 1:100; p = 0.027 for WF-3/Luc:splenocyte = 1:50; and p = 0.039 for WF-3/Luc:splenocyte = 1:10, Kruskal-Wallis test). The splenocytes treated with anti-BTLA Ab could generate higher tumor killing activities than those without anti-BTLA Ab. (5 mice in each group) f Bar figures of concentrations of various cytokines in ascites of various groups. Note : F1: IL-12; F2: TNF-α; F3: IFN-γ; F4: IL-6; F5: IL-10; and F6: TGF-β. The pro-inflammatory cytokines such as IL-12 ( p = 0.002), TNF-α ( p = 0.002), and IFN-γ ( p = 0.001) were highest with the chemotherapy combined with anti-BTLA Ab 20 μg/mouse. The concentrations of ant-inflammatory cytokines such as IL-6 ( p = 0.83), IL-10 ( p = 0.85), and TGF-β ( p = 0.84) did not show differences among the various groups (all statistical analyses by Kruskal-Wallis test). (5 mice in each group)

    Journal: Journal for Immunotherapy of Cancer

    Article Title: BTLA blockade enhances Cancer therapy by inhibiting IL-6/IL-10-induced CD19high B lymphocytes

    doi: 10.1186/s40425-019-0744-4

    Figure Lengend Snippet: Immunologic alterations in tumor-bearing mice treated with chemotherapy and/or anti-BTLA Ab. a The percentages of CD223 expression of CD4 + T lymphocytes in splenocytes of various therapeutic groups. The percentage of CD223 + CD4 + T lymphocytes in splenocytes was highest in paclitaxel combined with anti-BTLA Ab 20 μg/mouse group ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) b The percentages of CD223 expression of CD8 + T lymphocytes in splenocytes of various therapeutic groups. The percentage of CD223 + CD8 + T lymphocytes in splenocytes was also highest with paclitaxel combined with anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) c The percentages of CD223 expression of CD4 + T lymphocytes in TACs of ascites of various therapeutic groups. The percentage of CD223 + CD4 + T lymphocytes in TACs of ascites was highest with paclitaxel and anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) d The percentages of CD223 expression of CD8 + T lymphocytes in TACs of ascites of various therapeutic groups. The percentage of CD223 + CD8 + T lymphocytes in TACs of ascites was also highest with paclitaxel combined with anti-BTLA Ab 20 μg/mouse ( p = 0.001, Kruskal-Wallis test). (5 mice in each group) e Tumoricidal activity of splenocytes of tumor-bearing mice receiving chemotherapy treated with/without anti-BTLA Ab in vitro. e1 Representative luminescence figures of the in vitro tumor killing abilities of splenocytes by the IVIS system. (5 mice in each group) e2 Quantification of luminescence of in vitro tumor killing abilities of splenocytes by the IVIS system. Compared with the luminescence of WF-3/Luc cells co-cultured with splenocytes without anti-BTLA Ab, less luminal activity was detected in WF-3/Luc cells co-cultured with splenocytes receiving in vitro anti-BTLA Ab ( p = 0.021 for WF-3/Luc:splenocyte = 1:100; p = 0.027 for WF-3/Luc:splenocyte = 1:50; and p = 0.039 for WF-3/Luc:splenocyte = 1:10, Kruskal-Wallis test). The splenocytes treated with anti-BTLA Ab could generate higher tumor killing activities than those without anti-BTLA Ab. (5 mice in each group) f Bar figures of concentrations of various cytokines in ascites of various groups. Note : F1: IL-12; F2: TNF-α; F3: IFN-γ; F4: IL-6; F5: IL-10; and F6: TGF-β. The pro-inflammatory cytokines such as IL-12 ( p = 0.002), TNF-α ( p = 0.002), and IFN-γ ( p = 0.001) were highest with the chemotherapy combined with anti-BTLA Ab 20 μg/mouse. The concentrations of ant-inflammatory cytokines such as IL-6 ( p = 0.83), IL-10 ( p = 0.85), and TGF-β ( p = 0.84) did not show differences among the various groups (all statistical analyses by Kruskal-Wallis test). (5 mice in each group)

    Article Snippet: PBS, recombinant mouse IL-6 (20 ng/mL), IL-10 (20 ng/mL), or TGF-β (10 ng/mL) (PeproTech) was loaded with these collected B lymphocytes for 24 h. Then, the percentage of BTLA+ CD19hi B lymphocytes was analyzed by flow cytometry.

    Techniques: Mouse Assay, Expressing, Activity Assay, In Vitro, Cell Culture

    IL-6 and IL-10 could induce more BTLA + CD19 hi B lymphocytes through the AKT and STAT3 signaling pathways. a Representative figures of flow cytometric analyses of the expression of BTLA on various kinds of immunocytes of splenocytes. Note: A1: T lymphocytes; A2: NK cells; A3: B lymphocytes; A4: subgroups of B lymphocytes (zone 1: BTLA − CD19 hi ; zone 2: BTLA + CD19 hi ; zone 3: BTLA + CD19 low(lo) ; zone 4: BTLA + CD19 lo ). B lymphocytes, especially CD19 hi B lymphocytes, had higher percentages expressing the BTLA molecule. (5 mice in this analysis) b Kinetic alterations in BTLA + CD19 hi B lymphocytes in splenocytes of tumor-bearing mice after different days of tumor challenge. b1 Representative flow cytometric figures of percentages of BTLA + CD19 hi B lymphocytes in splenocytes on indicated days. (5 mice in each group) b2 Bar figures exhibited the percentages of BTLA + CD19 hi B lymphocytes in splenocytes on day 14 or day 35 after tumor challenge. The percentages of BTLA + CD19 hi B lymphocytes were higher on day 35 (17.74 ± 0.71%) than on day 14 (11.76 ± 0.52%) ( p = 0.009, Kruskal-Wallis test). (5 mice in each group) c Kinetic alterations in BTLA + CD19 hi B lymphocytes in TACs of ascites from tumor-bearing mice after different days of tumor challenge. c1 Representative flow cytometric figures of BTLA + CD19 hi B lymphocytes in TACs at indicated intervals. (5 mice in each group) c2 Bar figures of the percentages of BTLA + CD19 hi B lymphocytes in TACs on day 14 or day 35 after tumor challenge. The percentages of BTLA + CD19 hi B lymphocytes were higher on day 35 (48.94 ± 0.92%) than on day 14 (19.34 ± 0.88%) ( p = 0.007, Kruskal-Wallis test). (5 mice in each group) d Alterations in the percentages of BTLA + CD19 hi B lymphocytes in sorted B lymphocytes treated with IL-6, IL-10, or TGF-β, analyzed by flow cytometry. d1 Representative flow cytometric figures of the percentages of BTLA + CD19 hi B lymphocytes in sorted B cells. (5 mice in each group) d2 Bar figures of the percentages of BTLA + CD19 hi B lymphocytes in sorted B cells treated with respective cytokines. The percentages of BTLA + CD19 hi B lymphocytes increased under treatment with IL-6 or IL-10 compared with TGF-β ( p = 0.033, Kruskal-Wallis test). (5 mice in each group) e Various signaling molecules of sorted B lymphocytes treated with IL-6 and IL-10, detected by western blotting and flow cytometric analyses. e1 IL-6 (10 or 20 ng/mL) could stimulate phosphorylation of STAT3 and AKT in sorted B lymphocytes. (5 mice in each group) e2 Phosphorylation of STAT3 and AKT in sorted B cells also could be promoted by IL-10 (10 or 20 ng/mL). (5 mice in each group) e3 The inhibition of p-AKT by LY294002 showed inhibition of p-STAT3 (Lanes 3 and 9). However, the inhibition of p-STAT3 by BP-1-102 did not block activation of p-AKT (Lanes 4 and 10). Therefore, AKT activation was upstream of STAT3 in the IL-6/IL-10 signaling pathway. (5 mice in each group) e4 Percentages of BTLA + CD19 hi B lymphocytes in sorted B lymphocytes pretreated with respective Ab or specific inhibitor and then incubated with respective cytokine, analyzed by flow cytometry. The percentages of BTLA + CD19 hi B lymphocytes decreased when the B lymphocytes were pretreated with anti-BTLA Ab, LY294002 (AKT inhibitor), or BP-1-102 (STAT3 inhibitor) compared with PD98059 (ERK inhibitor). (5 mice in each group) f Anti-tumor effects of chemotherapy combined with various BTLA-related inhibitors. (F1) Diagrammatic representation of different treatment protocols using paclitaxel and various BTLA inhibitors. Note: Ga: paclitaxel 6 mg/kg; Gb: paclitaxel 6 mg/kg and LY294002 800 μg/mouse; Gc: paclitaxel 6 mg/kg and BP-1-102 40 μg/mouse; Gd: paclitaxel 6 mg/kg and anti-BTLA Ab 20 μg/mouse. (F2) Representative luminescence images of mice in various groups using the IVIS system on day 35 after tumor challenge. (5 mice in each group) (F3) Luminal analyses of tumor volumes in tumor-bearing mice with various regimens. Mice treated with paclitaxel and various BTLA-related inhibitors exhibited less luminescence than the paclitaxel-treated group ( p

    Journal: Journal for Immunotherapy of Cancer

    Article Title: BTLA blockade enhances Cancer therapy by inhibiting IL-6/IL-10-induced CD19high B lymphocytes

    doi: 10.1186/s40425-019-0744-4

    Figure Lengend Snippet: IL-6 and IL-10 could induce more BTLA + CD19 hi B lymphocytes through the AKT and STAT3 signaling pathways. a Representative figures of flow cytometric analyses of the expression of BTLA on various kinds of immunocytes of splenocytes. Note: A1: T lymphocytes; A2: NK cells; A3: B lymphocytes; A4: subgroups of B lymphocytes (zone 1: BTLA − CD19 hi ; zone 2: BTLA + CD19 hi ; zone 3: BTLA + CD19 low(lo) ; zone 4: BTLA + CD19 lo ). B lymphocytes, especially CD19 hi B lymphocytes, had higher percentages expressing the BTLA molecule. (5 mice in this analysis) b Kinetic alterations in BTLA + CD19 hi B lymphocytes in splenocytes of tumor-bearing mice after different days of tumor challenge. b1 Representative flow cytometric figures of percentages of BTLA + CD19 hi B lymphocytes in splenocytes on indicated days. (5 mice in each group) b2 Bar figures exhibited the percentages of BTLA + CD19 hi B lymphocytes in splenocytes on day 14 or day 35 after tumor challenge. The percentages of BTLA + CD19 hi B lymphocytes were higher on day 35 (17.74 ± 0.71%) than on day 14 (11.76 ± 0.52%) ( p = 0.009, Kruskal-Wallis test). (5 mice in each group) c Kinetic alterations in BTLA + CD19 hi B lymphocytes in TACs of ascites from tumor-bearing mice after different days of tumor challenge. c1 Representative flow cytometric figures of BTLA + CD19 hi B lymphocytes in TACs at indicated intervals. (5 mice in each group) c2 Bar figures of the percentages of BTLA + CD19 hi B lymphocytes in TACs on day 14 or day 35 after tumor challenge. The percentages of BTLA + CD19 hi B lymphocytes were higher on day 35 (48.94 ± 0.92%) than on day 14 (19.34 ± 0.88%) ( p = 0.007, Kruskal-Wallis test). (5 mice in each group) d Alterations in the percentages of BTLA + CD19 hi B lymphocytes in sorted B lymphocytes treated with IL-6, IL-10, or TGF-β, analyzed by flow cytometry. d1 Representative flow cytometric figures of the percentages of BTLA + CD19 hi B lymphocytes in sorted B cells. (5 mice in each group) d2 Bar figures of the percentages of BTLA + CD19 hi B lymphocytes in sorted B cells treated with respective cytokines. The percentages of BTLA + CD19 hi B lymphocytes increased under treatment with IL-6 or IL-10 compared with TGF-β ( p = 0.033, Kruskal-Wallis test). (5 mice in each group) e Various signaling molecules of sorted B lymphocytes treated with IL-6 and IL-10, detected by western blotting and flow cytometric analyses. e1 IL-6 (10 or 20 ng/mL) could stimulate phosphorylation of STAT3 and AKT in sorted B lymphocytes. (5 mice in each group) e2 Phosphorylation of STAT3 and AKT in sorted B cells also could be promoted by IL-10 (10 or 20 ng/mL). (5 mice in each group) e3 The inhibition of p-AKT by LY294002 showed inhibition of p-STAT3 (Lanes 3 and 9). However, the inhibition of p-STAT3 by BP-1-102 did not block activation of p-AKT (Lanes 4 and 10). Therefore, AKT activation was upstream of STAT3 in the IL-6/IL-10 signaling pathway. (5 mice in each group) e4 Percentages of BTLA + CD19 hi B lymphocytes in sorted B lymphocytes pretreated with respective Ab or specific inhibitor and then incubated with respective cytokine, analyzed by flow cytometry. The percentages of BTLA + CD19 hi B lymphocytes decreased when the B lymphocytes were pretreated with anti-BTLA Ab, LY294002 (AKT inhibitor), or BP-1-102 (STAT3 inhibitor) compared with PD98059 (ERK inhibitor). (5 mice in each group) f Anti-tumor effects of chemotherapy combined with various BTLA-related inhibitors. (F1) Diagrammatic representation of different treatment protocols using paclitaxel and various BTLA inhibitors. Note: Ga: paclitaxel 6 mg/kg; Gb: paclitaxel 6 mg/kg and LY294002 800 μg/mouse; Gc: paclitaxel 6 mg/kg and BP-1-102 40 μg/mouse; Gd: paclitaxel 6 mg/kg and anti-BTLA Ab 20 μg/mouse. (F2) Representative luminescence images of mice in various groups using the IVIS system on day 35 after tumor challenge. (5 mice in each group) (F3) Luminal analyses of tumor volumes in tumor-bearing mice with various regimens. Mice treated with paclitaxel and various BTLA-related inhibitors exhibited less luminescence than the paclitaxel-treated group ( p

    Article Snippet: PBS, recombinant mouse IL-6 (20 ng/mL), IL-10 (20 ng/mL), or TGF-β (10 ng/mL) (PeproTech) was loaded with these collected B lymphocytes for 24 h. Then, the percentage of BTLA+ CD19hi B lymphocytes was analyzed by flow cytometry.

    Techniques: Flow Cytometry, Expressing, Mouse Assay, Cytometry, Western Blot, Inhibition, Blocking Assay, Activation Assay, Incubation

    TGFβ, IL-6 and IL-23 synergistically up-regulate IL-17 production during collagen re-stimulation, but IL-4 and IFNγ continue to suppress

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

    Article Title: Sensitivity and resistance to regulation by IL-4 during Th17 maturation

    doi: 10.4049/jimmunol.1002860

    Figure Lengend Snippet: TGFβ, IL-6 and IL-23 synergistically up-regulate IL-17 production during collagen re-stimulation, but IL-4 and IFNγ continue to suppress

    Article Snippet: BM-DCs and naïve T cells were plated in 6 well plates in basic RPMI at 0.125×106 BM-DCs and 0.25×106 naïve T cells per mL with 4 µg/mL anti-CD3 (145-2C11), 10 µg/mL anti-IL-4 (11B11), 10 µg/mL anti-IFNγ (R4-6A2), 1 ng/mL recombinant human TGF-β1 (Peprotech), 20ng/mL recombinant mouse IL-6 (Peprotech) and 10 ng/mL recombinant mouse IL-23 (eBioscience).

    Techniques:

    MKP-1 signaling regulates Arid5a mRNA stability by inducing the translocalization of AUF-1 from the nucleus to the cytoplasm. ( A and B ) Analysis of MKP-1 mRNA and protein levels in MEF cells that were transfected with control siRNA and MKP-1 siRNA. ( C ) Quantitative real-time PCR analysis of AUF-1 mRNA levels in control and MKP-1 knockdown cells that were stimulated with LPS (3 μg/ml). ( D ) AUF-1 (p45, p42, p40, p37) levels in the cytoplasm versus the nucleus in control and MKP-1 knockdown MEF cells that were stimulated using LPS for 0–4 h. β-tubulin and Lamin A/C were used as the loading controls for cytoplasmic proteins and nuclear proteins, respectively. ( E ) Quantitative real-time PCR analysis of mRNA levels of Arid5a in control and MKP-1 knockdown MEFs that were stimulated with LPS and then treated with Actinomycin D for 0–90 min. ( F and G ) Quantitative real-time PCR analysis of mRNA levels of Arid5a and IL-6 in control and MKP-1 knockdown MEFs that were stimulated with LPS. ( H ) Schematic diagram illustrating the post-transcriptional regulation of Arid5a mRNA. LPS signaling induces MKP-1, which in turn signals AUF-1 to translocate from the nucleus to the cytoplasm. In the cytoplasm, AUF-1 binds to AU-rich elements on the Arid5a 3΄ UTR to destabilize the Arid5a mRNA. This results in the inhibition of the stabilization of the IL-6 mRNA. All data are shown as the mean ± SD of three independent experiments. N.S., not significant. * P

    Journal: Nucleic Acids Research

    Article Title: TLR4-induced NF-κB and MAPK signaling regulate the IL-6 mRNA stabilizing protein Arid5a

    doi: 10.1093/nar/gkx064

    Figure Lengend Snippet: MKP-1 signaling regulates Arid5a mRNA stability by inducing the translocalization of AUF-1 from the nucleus to the cytoplasm. ( A and B ) Analysis of MKP-1 mRNA and protein levels in MEF cells that were transfected with control siRNA and MKP-1 siRNA. ( C ) Quantitative real-time PCR analysis of AUF-1 mRNA levels in control and MKP-1 knockdown cells that were stimulated with LPS (3 μg/ml). ( D ) AUF-1 (p45, p42, p40, p37) levels in the cytoplasm versus the nucleus in control and MKP-1 knockdown MEF cells that were stimulated using LPS for 0–4 h. β-tubulin and Lamin A/C were used as the loading controls for cytoplasmic proteins and nuclear proteins, respectively. ( E ) Quantitative real-time PCR analysis of mRNA levels of Arid5a in control and MKP-1 knockdown MEFs that were stimulated with LPS and then treated with Actinomycin D for 0–90 min. ( F and G ) Quantitative real-time PCR analysis of mRNA levels of Arid5a and IL-6 in control and MKP-1 knockdown MEFs that were stimulated with LPS. ( H ) Schematic diagram illustrating the post-transcriptional regulation of Arid5a mRNA. LPS signaling induces MKP-1, which in turn signals AUF-1 to translocate from the nucleus to the cytoplasm. In the cytoplasm, AUF-1 binds to AU-rich elements on the Arid5a 3΄ UTR to destabilize the Arid5a mRNA. This results in the inhibition of the stabilization of the IL-6 mRNA. All data are shown as the mean ± SD of three independent experiments. N.S., not significant. * P

    Article Snippet: Recombinant murine IL-6 was obtained from Peprotech, MG-132 (proteasome inhibitor) was obtained from Calbiochem, SB203580 (p38 MAP kinase inhibitor) was obtained from Abcam and Stat3 Inhibitor VI (S3I-201) was obtained from Santa Cruz Biotechnology.

    Techniques: Transfection, Real-time Polymerase Chain Reaction, Inhibition

    AUF-1 destabilizes the Arid5a mRNA by recognizing the Arid5a 3΄ UTR. ( A ) Identification of the RNA-binding protein AUF-1 on the Arid5a 3΄ UTR. Peritoneal macrophages were stimulated with or without LPS (1 μg/ml) for 4 h. Whole-cell lysates were then obtained and mixed with binding protein G beads in solution with the Arid5a 3΄ UTR (containing BrdU), and the eluted buffer was then subjected to SDS-PAGE. The gel that was run using the complex containing the Arid5a 3΄ UTR and beads was stained using CBB after SDS-PAGE and then separated into five compartments that were independently analyzed using LC-MS/MS. ( B and C ) Analysis of AUF-1 mRNA and protein levels in MEF cells that were transfected with control siRNA or AUF-1 siRNA. ( D–F ) Arid5a mRNA (D) and protein expression (E) and IL-6 mRNA expression (F) in the control and AUF-1 knockdown MEF cells at 4 h after stimulation with LPS (5 μg/ml). ( G ) Diagram of the region of the pGL3 vector that encodes the Arid5a 3΄ UTR (1–3532), Arid5a 3΄ UTR (1-800), or Arid5a 3΄ UTR (800–3532). The black bar shows the locations of AU-rich elements (AREs). ( H–J ) The luciferase activities of each of the vectors that encodes one of three different fragments of the Arid5a 3΄ UTR are shown in G. The results were obtained using MEF cells that were transfected for 48 h with an AUF-1 expression vector or an empty vector. ( K ) Quantitative real-time PCR analysis of the Arid5a mRNA in the control and AUF-1 knockdown MEFs that were stimulated for 2 h with LPS (3 μg/ml). The cells were then treated for 0–60 min with Actinomycin D. ( L ) A predicted ARE region in the Arid5a 3΄ UTR is shown in red. ( M ) Electrophoretic mobility shift assay (EMSA) demonstrating the interaction between the AUF-1 recombinant protein and 3΄-biotinylated fragments with the Arid5a 3΄ UTR, as shown in L. ( N ) Putative binding mode of mouse AUF-1 protein with the homology model of AUF-1 shown as a cartoon with rainbow colors from N (blue) and to C (red). The AUUUA RNA fragment from PDB entry 1g2e is shown as an orange cartoon with light green residues in stick representation and with the flexibly docked RNA models shown as orange backbone traces. All data are shown as the mean ± SD of three independent experiments. Error bars indicate the mean ± SD. * P

    Journal: Nucleic Acids Research

    Article Title: TLR4-induced NF-κB and MAPK signaling regulate the IL-6 mRNA stabilizing protein Arid5a

    doi: 10.1093/nar/gkx064

    Figure Lengend Snippet: AUF-1 destabilizes the Arid5a mRNA by recognizing the Arid5a 3΄ UTR. ( A ) Identification of the RNA-binding protein AUF-1 on the Arid5a 3΄ UTR. Peritoneal macrophages were stimulated with or without LPS (1 μg/ml) for 4 h. Whole-cell lysates were then obtained and mixed with binding protein G beads in solution with the Arid5a 3΄ UTR (containing BrdU), and the eluted buffer was then subjected to SDS-PAGE. The gel that was run using the complex containing the Arid5a 3΄ UTR and beads was stained using CBB after SDS-PAGE and then separated into five compartments that were independently analyzed using LC-MS/MS. ( B and C ) Analysis of AUF-1 mRNA and protein levels in MEF cells that were transfected with control siRNA or AUF-1 siRNA. ( D–F ) Arid5a mRNA (D) and protein expression (E) and IL-6 mRNA expression (F) in the control and AUF-1 knockdown MEF cells at 4 h after stimulation with LPS (5 μg/ml). ( G ) Diagram of the region of the pGL3 vector that encodes the Arid5a 3΄ UTR (1–3532), Arid5a 3΄ UTR (1-800), or Arid5a 3΄ UTR (800–3532). The black bar shows the locations of AU-rich elements (AREs). ( H–J ) The luciferase activities of each of the vectors that encodes one of three different fragments of the Arid5a 3΄ UTR are shown in G. The results were obtained using MEF cells that were transfected for 48 h with an AUF-1 expression vector or an empty vector. ( K ) Quantitative real-time PCR analysis of the Arid5a mRNA in the control and AUF-1 knockdown MEFs that were stimulated for 2 h with LPS (3 μg/ml). The cells were then treated for 0–60 min with Actinomycin D. ( L ) A predicted ARE region in the Arid5a 3΄ UTR is shown in red. ( M ) Electrophoretic mobility shift assay (EMSA) demonstrating the interaction between the AUF-1 recombinant protein and 3΄-biotinylated fragments with the Arid5a 3΄ UTR, as shown in L. ( N ) Putative binding mode of mouse AUF-1 protein with the homology model of AUF-1 shown as a cartoon with rainbow colors from N (blue) and to C (red). The AUUUA RNA fragment from PDB entry 1g2e is shown as an orange cartoon with light green residues in stick representation and with the flexibly docked RNA models shown as orange backbone traces. All data are shown as the mean ± SD of three independent experiments. Error bars indicate the mean ± SD. * P

    Article Snippet: Recombinant murine IL-6 was obtained from Peprotech, MG-132 (proteasome inhibitor) was obtained from Calbiochem, SB203580 (p38 MAP kinase inhibitor) was obtained from Abcam and Stat3 Inhibitor VI (S3I-201) was obtained from Santa Cruz Biotechnology.

    Techniques: RNA Binding Assay, Binding Assay, SDS Page, Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Transfection, Expressing, Plasmid Preparation, Luciferase, Real-time Polymerase Chain Reaction, Electrophoretic Mobility Shift Assay, Recombinant

    IL-6 stimulates the expression of Arid5a via STAT3 in response to LPS. ( A ) Arid5a mRNA expression was measured using qPCR in MEFs stimulated with IL-6 (35 ng/ml) for 1 h. ( B ) IL-6 mRNA expression was knocked down in MEF cells using an IL-6 siRNA followed by stimulation with LPS (10 μg/ml). ( C ) Expression of Arid5a in LPS-stimulated knockdown MEF cells (10 μg/ml) for 1 h. ( D ) Expression of Arid5a mRNA in MEFs that were pretreated with a Stat3 inhibitor (50 μM) or DMSO (control) for 45 min and then stimulated with IL-6 (35 ng/ml) for 1 h. ( E ) Immunoblot assays of Stat3 expression in MEFs transfected with 20 nM control or Stat3 siRNA. ( F ) Arid5a mRNA expression in the control and Stat3 knockdown MEFs after stimulation with IL-6 (35 ng/ml). ( G ) Expression of Arid5a mRNA in MEFs that were pretreated with a Stat3 inhibitor (50 μM) or DMSO (control) for 45 min and then stimulated with LPS (10 μg/mL) for 1 h. ( H ) Luciferase activity of the Arid5a promoter was determined as described above in 1E in combination with the empty vector or WT Stat3 and its phosphorylation site mutants (Y705F and S727A) expression vectors. ( I ) ChIP assay was performed as described above in 1G using LPS stimulated-macrophages, and chromatins were immunoprecipitated with anti-Stat3 antibody and anti-IgG (negative control). ( J ) Schematic diagram illustrating the mechanism proposed in this study, in which the TLR4/IKK/NF-κB and IL-6/STAT3 signaling pathways transcriptionally regulate Arid5a gene expression. During LPS signaling, NF-κB upregulates Arid5a, which in turn increases the production of IL-6 by stabilizing its mRNA. IL-6 further stimulates the expression of Arid5a and IL-6 via a positive feedback loop. The values are shown relative to normalized levels after cells were transfected using an empty vector. All data are shown the mean ± SD of three independent experiments. Error bars indicate the mean ± SD. NS, not significant; * P

    Journal: Nucleic Acids Research

    Article Title: TLR4-induced NF-κB and MAPK signaling regulate the IL-6 mRNA stabilizing protein Arid5a

    doi: 10.1093/nar/gkx064

    Figure Lengend Snippet: IL-6 stimulates the expression of Arid5a via STAT3 in response to LPS. ( A ) Arid5a mRNA expression was measured using qPCR in MEFs stimulated with IL-6 (35 ng/ml) for 1 h. ( B ) IL-6 mRNA expression was knocked down in MEF cells using an IL-6 siRNA followed by stimulation with LPS (10 μg/ml). ( C ) Expression of Arid5a in LPS-stimulated knockdown MEF cells (10 μg/ml) for 1 h. ( D ) Expression of Arid5a mRNA in MEFs that were pretreated with a Stat3 inhibitor (50 μM) or DMSO (control) for 45 min and then stimulated with IL-6 (35 ng/ml) for 1 h. ( E ) Immunoblot assays of Stat3 expression in MEFs transfected with 20 nM control or Stat3 siRNA. ( F ) Arid5a mRNA expression in the control and Stat3 knockdown MEFs after stimulation with IL-6 (35 ng/ml). ( G ) Expression of Arid5a mRNA in MEFs that were pretreated with a Stat3 inhibitor (50 μM) or DMSO (control) for 45 min and then stimulated with LPS (10 μg/mL) for 1 h. ( H ) Luciferase activity of the Arid5a promoter was determined as described above in 1E in combination with the empty vector or WT Stat3 and its phosphorylation site mutants (Y705F and S727A) expression vectors. ( I ) ChIP assay was performed as described above in 1G using LPS stimulated-macrophages, and chromatins were immunoprecipitated with anti-Stat3 antibody and anti-IgG (negative control). ( J ) Schematic diagram illustrating the mechanism proposed in this study, in which the TLR4/IKK/NF-κB and IL-6/STAT3 signaling pathways transcriptionally regulate Arid5a gene expression. During LPS signaling, NF-κB upregulates Arid5a, which in turn increases the production of IL-6 by stabilizing its mRNA. IL-6 further stimulates the expression of Arid5a and IL-6 via a positive feedback loop. The values are shown relative to normalized levels after cells were transfected using an empty vector. All data are shown the mean ± SD of three independent experiments. Error bars indicate the mean ± SD. NS, not significant; * P

    Article Snippet: Recombinant murine IL-6 was obtained from Peprotech, MG-132 (proteasome inhibitor) was obtained from Calbiochem, SB203580 (p38 MAP kinase inhibitor) was obtained from Abcam and Stat3 Inhibitor VI (S3I-201) was obtained from Santa Cruz Biotechnology.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Luciferase, Activity Assay, Plasmid Preparation, Chromatin Immunoprecipitation, Immunoprecipitation, Negative Control

    WWP1 E3 ubiquitin ligase mediates Arid5a polyubiquitination. ( A ) Analysis of WWP1 mRNA expression in MEF cells that were transfected with control siRNA and WWP1 siRNA (20 nM each). ( B ) MEF cells were treated with 20 nM control and E3 ubiquitin ligase siRNAs (WWP1, Atrogin1, Trim29, Trim38 and Smurf1) for 72 h and transfected to express hemagglutinin-tagged ubiquitin (HA-Ub) and Flag-tagged Arid5a (as indicated above the lanes). The cells were then stimulated for 6 h with LPS (10 μg/ml) and incubated for 6 h with MG-132 (1 μM) followed by immunoprecipitation with anti-Flag antibodies and immunoblot analyses of ubiquitin (using anti-HA) and Arid5a (using anti-Flag). Below, immunoblots of whole-cell lysates treated with anti-Flag antibody. ( C and D ) Immunoblot analysis of molecular interactions in HEK293TLR4 cells following transfection for 48 h with WT Myc-Arid5a, Flag-WWP1 (C) and HA-p38, Flag-WWP1 (D). The lysates were immunoprecipitated using anti-Myc (C) and anti-Flag (D) antibodies. This was followed by an immunoblot analysis of WWP1 (with anti-Flag antibody), p38 (with anti-HA antibody). Below, immunoblots of whole-cell lysates that were treated with anti-Myc, anti-Flag (C) and anti-HA, anti-Flag (D) antibodies. ( E ) Immunoassays of HEK293TLR4 cells transfected for 48 h to express HA-tagged Ub, Myc-tagged Arid5a or Flag-tagged WWP1 expression vector (as indicated above the lanes), then incubated for 6 h with MG-132 (1 μM) followed by immunoprecipitation with anti-Myc antibodies and immunoblot analyses of ubiquitin (using anti-HA), Below, immunoblots of whole-cell lysates treated with anti-Flag or anti-Myc antibodies. ( F ) IL-6 protein levels in the supernatants of control and WWP1 knockdown MEF cells at 24 h after stimulation with LPS (10 μg/ml). The data are representative of two–three independent experiments.

    Journal: Nucleic Acids Research

    Article Title: TLR4-induced NF-κB and MAPK signaling regulate the IL-6 mRNA stabilizing protein Arid5a

    doi: 10.1093/nar/gkx064

    Figure Lengend Snippet: WWP1 E3 ubiquitin ligase mediates Arid5a polyubiquitination. ( A ) Analysis of WWP1 mRNA expression in MEF cells that were transfected with control siRNA and WWP1 siRNA (20 nM each). ( B ) MEF cells were treated with 20 nM control and E3 ubiquitin ligase siRNAs (WWP1, Atrogin1, Trim29, Trim38 and Smurf1) for 72 h and transfected to express hemagglutinin-tagged ubiquitin (HA-Ub) and Flag-tagged Arid5a (as indicated above the lanes). The cells were then stimulated for 6 h with LPS (10 μg/ml) and incubated for 6 h with MG-132 (1 μM) followed by immunoprecipitation with anti-Flag antibodies and immunoblot analyses of ubiquitin (using anti-HA) and Arid5a (using anti-Flag). Below, immunoblots of whole-cell lysates treated with anti-Flag antibody. ( C and D ) Immunoblot analysis of molecular interactions in HEK293TLR4 cells following transfection for 48 h with WT Myc-Arid5a, Flag-WWP1 (C) and HA-p38, Flag-WWP1 (D). The lysates were immunoprecipitated using anti-Myc (C) and anti-Flag (D) antibodies. This was followed by an immunoblot analysis of WWP1 (with anti-Flag antibody), p38 (with anti-HA antibody). Below, immunoblots of whole-cell lysates that were treated with anti-Myc, anti-Flag (C) and anti-HA, anti-Flag (D) antibodies. ( E ) Immunoassays of HEK293TLR4 cells transfected for 48 h to express HA-tagged Ub, Myc-tagged Arid5a or Flag-tagged WWP1 expression vector (as indicated above the lanes), then incubated for 6 h with MG-132 (1 μM) followed by immunoprecipitation with anti-Myc antibodies and immunoblot analyses of ubiquitin (using anti-HA), Below, immunoblots of whole-cell lysates treated with anti-Flag or anti-Myc antibodies. ( F ) IL-6 protein levels in the supernatants of control and WWP1 knockdown MEF cells at 24 h after stimulation with LPS (10 μg/ml). The data are representative of two–three independent experiments.

    Article Snippet: Recombinant murine IL-6 was obtained from Peprotech, MG-132 (proteasome inhibitor) was obtained from Calbiochem, SB203580 (p38 MAP kinase inhibitor) was obtained from Abcam and Stat3 Inhibitor VI (S3I-201) was obtained from Santa Cruz Biotechnology.

    Techniques: Expressing, Transfection, Incubation, Immunoprecipitation, Western Blot, Plasmid Preparation

    The phosphorylation-mediated degradation of Arid5a affects IL-6 mRNA expression. ( A ) Immunoblot analysis of lysates obtained from Arid5a-deficient MEF cells that expressed an empty vector (EV) or an expression vector containing WT Flag-tagged Arid5a or S253A, S433A and S458A-mutant Flag-Arid5a. The cells were stimulated for 0–12 h (above lane) with LPS (5 μg/ml) and then probed with anti-Flag or anti-GAPDH antibodies. ( B ) Densitometry analysis with results presented as the ratio to GAPDH. ( C ) Immunoblot assay of p38 expression in MEF cells transfected with 20 nM control or p38 siRNA. ( D and E ) Immunoblot (D) and densitometry analysis (E) of Flag-tagged Arid5a protein expression following LPS stimulation in control and p38 knockdown MEFs as described above in (A and B). ( F ) Immunoassay of HEK293TLR4 cells transfected for 48 h to express WT HA-tagged ubiquitin (HA-Ub), K48-HA-Ub, Myc-tagged Arid5a or p38 expression vector (as indicated above the lanes), then incubated for 6 h with MG-132 (1 μM) followed by immunoprecipitation with anti-Myc antibodies and immunoblot analyses of ubiquitin (using anti-HA), p38 (using anti-p38) and Arid5a (using anti-Myc). Below, immunoblots of whole-cell lysates treated with anti-p38 or anti-Myc antibodies. ( G ) Quantitative PCR analysis of the expression level of IL-6 mRNA in total RNA obtained from the cells described in A and presented relative to the level of GAPDH RNA. ( H ) Schematic diagram representing the regulatory mechanism proposed by which degradation of Arid5a by p38 signaling regulates the stability of IL-6 mRNA. The activation of p38 by LPS resulted in the phosphorylation of Arid5a at serine residues 253, 433 and 458, which leads to its ubiquitination on lysine residues 80 and 89 and its subsequent degradation. The degradation of Arid5a results in the inhibition of the stabilization of IL-6 mRNA. Mutating Arid5a at the above described phosphorylation sites blocked it from being degraded, which resulted in the overproduction of IL-6 because its mRNA remained stabilized. The cross shown in green indicates the phosphorylation sites that were mutated in Arid5a, and the cross shown in red indicates its inhibited function. The data are representative of two–three independent experiments. Error bars indicate the mean ± SD. ** P

    Journal: Nucleic Acids Research

    Article Title: TLR4-induced NF-κB and MAPK signaling regulate the IL-6 mRNA stabilizing protein Arid5a

    doi: 10.1093/nar/gkx064

    Figure Lengend Snippet: The phosphorylation-mediated degradation of Arid5a affects IL-6 mRNA expression. ( A ) Immunoblot analysis of lysates obtained from Arid5a-deficient MEF cells that expressed an empty vector (EV) or an expression vector containing WT Flag-tagged Arid5a or S253A, S433A and S458A-mutant Flag-Arid5a. The cells were stimulated for 0–12 h (above lane) with LPS (5 μg/ml) and then probed with anti-Flag or anti-GAPDH antibodies. ( B ) Densitometry analysis with results presented as the ratio to GAPDH. ( C ) Immunoblot assay of p38 expression in MEF cells transfected with 20 nM control or p38 siRNA. ( D and E ) Immunoblot (D) and densitometry analysis (E) of Flag-tagged Arid5a protein expression following LPS stimulation in control and p38 knockdown MEFs as described above in (A and B). ( F ) Immunoassay of HEK293TLR4 cells transfected for 48 h to express WT HA-tagged ubiquitin (HA-Ub), K48-HA-Ub, Myc-tagged Arid5a or p38 expression vector (as indicated above the lanes), then incubated for 6 h with MG-132 (1 μM) followed by immunoprecipitation with anti-Myc antibodies and immunoblot analyses of ubiquitin (using anti-HA), p38 (using anti-p38) and Arid5a (using anti-Myc). Below, immunoblots of whole-cell lysates treated with anti-p38 or anti-Myc antibodies. ( G ) Quantitative PCR analysis of the expression level of IL-6 mRNA in total RNA obtained from the cells described in A and presented relative to the level of GAPDH RNA. ( H ) Schematic diagram representing the regulatory mechanism proposed by which degradation of Arid5a by p38 signaling regulates the stability of IL-6 mRNA. The activation of p38 by LPS resulted in the phosphorylation of Arid5a at serine residues 253, 433 and 458, which leads to its ubiquitination on lysine residues 80 and 89 and its subsequent degradation. The degradation of Arid5a results in the inhibition of the stabilization of IL-6 mRNA. Mutating Arid5a at the above described phosphorylation sites blocked it from being degraded, which resulted in the overproduction of IL-6 because its mRNA remained stabilized. The cross shown in green indicates the phosphorylation sites that were mutated in Arid5a, and the cross shown in red indicates its inhibited function. The data are representative of two–three independent experiments. Error bars indicate the mean ± SD. ** P

    Article Snippet: Recombinant murine IL-6 was obtained from Peprotech, MG-132 (proteasome inhibitor) was obtained from Calbiochem, SB203580 (p38 MAP kinase inhibitor) was obtained from Abcam and Stat3 Inhibitor VI (S3I-201) was obtained from Santa Cruz Biotechnology.

    Techniques: Expressing, Plasmid Preparation, Mutagenesis, Transfection, Incubation, Immunoprecipitation, Western Blot, Real-time Polymerase Chain Reaction, Activation Assay, Inhibition

    Vagal signals increase macrophage IL-6 production after PHx. a Relative gene expressions of Il-6 after surgery in PHx-sham mice ( n = 4 per group) and PHx-HV mice ( n = 4 per group). b Relative gene expressions of Il-6 after PHx-sham in control liposome-treated ( n = 5–6 per group) and clodronate liposome-treated mice ( n = 5–6 per group), and after PHx-HV in clodronate liposome-treated mice ( n = 5–6 per group). c Relative gene expression of Il-6 in primary macrophages after 4 h of stimulation with vehicle ( n = 6), 100 µM carbachol ( n = 6), or both 100 µM carbachol and 100 µM atropine ( n = 6). d Relative gene expressions of Il-6 after PHx in vehicle-treated ( n = 4 per group) and atropine-treated ( n = 4 per group) mice. * P

    Journal: Nature Communications

    Article Title: Vagus-macrophage-hepatocyte link promotes post-injury liver regeneration and whole-body survival through hepatic FoxM1 activation

    doi: 10.1038/s41467-018-07747-0

    Figure Lengend Snippet: Vagal signals increase macrophage IL-6 production after PHx. a Relative gene expressions of Il-6 after surgery in PHx-sham mice ( n = 4 per group) and PHx-HV mice ( n = 4 per group). b Relative gene expressions of Il-6 after PHx-sham in control liposome-treated ( n = 5–6 per group) and clodronate liposome-treated mice ( n = 5–6 per group), and after PHx-HV in clodronate liposome-treated mice ( n = 5–6 per group). c Relative gene expression of Il-6 in primary macrophages after 4 h of stimulation with vehicle ( n = 6), 100 µM carbachol ( n = 6), or both 100 µM carbachol and 100 µM atropine ( n = 6). d Relative gene expressions of Il-6 after PHx in vehicle-treated ( n = 4 per group) and atropine-treated ( n = 4 per group) mice. * P

    Article Snippet: At 48 h after isolation, hepatocytes were incubated with 100 ng/ml recombinant murine IL-6 (PEPROTECH, London, UK) for 6 h, and then collected for RNA extraction.

    Techniques: Mouse Assay, Expressing

    IL-6 mediates vagal signal-induced hepatic FoxM1 activation. a Relative expressions of Foxm1 , its target genes, and MKi67 in primary hepatocytes isolated from control and iFoxM1LKO mice treated with 100 ng/ml IL-6 ( n = 7–8 per group) and vehicle ( n = 7–8 per group) for 6 h. b (Left panels) Representative images of liver extract immunoblottings with anti-phospho-STAT3, total STAT3, and actin. (Right panel) Relative intensities of phospho/total STAT3 in livers from sham operation for PHx (SO)- ( n = 6), PHx-sham- ( n = 6), and PHx-HV mice ( n = 6). c Relative expressions of Foxm1 , its target genes, and MKi67 in the liver 2 days after surgery from SO + IgG ( n = 5), PHx + IgG ( n = 5), and PHx + anti-IL-6 antibody- ( n = 5) treated mice. Data are presented as means ± SEM. * P

    Journal: Nature Communications

    Article Title: Vagus-macrophage-hepatocyte link promotes post-injury liver regeneration and whole-body survival through hepatic FoxM1 activation

    doi: 10.1038/s41467-018-07747-0

    Figure Lengend Snippet: IL-6 mediates vagal signal-induced hepatic FoxM1 activation. a Relative expressions of Foxm1 , its target genes, and MKi67 in primary hepatocytes isolated from control and iFoxM1LKO mice treated with 100 ng/ml IL-6 ( n = 7–8 per group) and vehicle ( n = 7–8 per group) for 6 h. b (Left panels) Representative images of liver extract immunoblottings with anti-phospho-STAT3, total STAT3, and actin. (Right panel) Relative intensities of phospho/total STAT3 in livers from sham operation for PHx (SO)- ( n = 6), PHx-sham- ( n = 6), and PHx-HV mice ( n = 6). c Relative expressions of Foxm1 , its target genes, and MKi67 in the liver 2 days after surgery from SO + IgG ( n = 5), PHx + IgG ( n = 5), and PHx + anti-IL-6 antibody- ( n = 5) treated mice. Data are presented as means ± SEM. * P

    Article Snippet: At 48 h after isolation, hepatocytes were incubated with 100 ng/ml recombinant murine IL-6 (PEPROTECH, London, UK) for 6 h, and then collected for RNA extraction.

    Techniques: Activation Assay, Isolation, Mouse Assay