Structured Review

BioLegend interleukin 6
Levels of proinflammatory cytokines released from S . Typhi- or S . Typhimurium-infected MDMs. Shown are levels of TNF-α (A) and <t>IL-6</t> (B) released from MDMs after 2.5 and 24 h of S . Typhi infection and TNF-α (C) and IL-6 (D) released from MDMs after 2.5 and 24 h of S . Typhimurium infection. Statistical significance was determined by Student's t test comparing cytokine levels between LPS modification mutant-infected samples and the WT-infected sample collected at that time point. *, P
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1) Product Images from "Cathelicidin Antimicrobial Peptide Expression Is Not Induced or Required for Bacterial Clearance during Salmonella enterica Infection of Human Monocyte-Derived Macrophages"

Article Title: Cathelicidin Antimicrobial Peptide Expression Is Not Induced or Required for Bacterial Clearance during Salmonella enterica Infection of Human Monocyte-Derived Macrophages

Journal: Infection and Immunity

doi: 10.1128/IAI.00672-12

Levels of proinflammatory cytokines released from S . Typhi- or S . Typhimurium-infected MDMs. Shown are levels of TNF-α (A) and IL-6 (B) released from MDMs after 2.5 and 24 h of S . Typhi infection and TNF-α (C) and IL-6 (D) released from MDMs after 2.5 and 24 h of S . Typhimurium infection. Statistical significance was determined by Student's t test comparing cytokine levels between LPS modification mutant-infected samples and the WT-infected sample collected at that time point. *, P
Figure Legend Snippet: Levels of proinflammatory cytokines released from S . Typhi- or S . Typhimurium-infected MDMs. Shown are levels of TNF-α (A) and IL-6 (B) released from MDMs after 2.5 and 24 h of S . Typhi infection and TNF-α (C) and IL-6 (D) released from MDMs after 2.5 and 24 h of S . Typhimurium infection. Statistical significance was determined by Student's t test comparing cytokine levels between LPS modification mutant-infected samples and the WT-infected sample collected at that time point. *, P

Techniques Used: Infection, Modification, Mutagenesis

2) Product Images from "A Low-Cost Mechanical Stretching Device for Uniaxial Strain of Cells: A Platform for Pedagogy in Mechanobiology"

Article Title: A Low-Cost Mechanical Stretching Device for Uniaxial Strain of Cells: A Platform for Pedagogy in Mechanobiology

Journal: Journal of Biomechanical Engineering

doi: 10.1115/1.4039949

Macrophage inflammatory cytokine secretion is unaffected by 10% cyclic uniaxial stretch. Secretion of TNF-α (left), IL-6 (middle), and MCP-1 (right) for unstimulated and IFN-γ/LPS stimulated macrophages subjected to either 0% or 10% cyclic uniaxial stretch. Data are normalized to IFN-γ/LPS stimulated and stretched condition. Error bars indicate standard deviation of the mean for three separate experiments.
Figure Legend Snippet: Macrophage inflammatory cytokine secretion is unaffected by 10% cyclic uniaxial stretch. Secretion of TNF-α (left), IL-6 (middle), and MCP-1 (right) for unstimulated and IFN-γ/LPS stimulated macrophages subjected to either 0% or 10% cyclic uniaxial stretch. Data are normalized to IFN-γ/LPS stimulated and stretched condition. Error bars indicate standard deviation of the mean for three separate experiments.

Techniques Used: Standard Deviation

3) Product Images from "LOX-1 Deletion Improves Neutrophil Responses, Enhances Bacterial Clearance, and Reduces Lung Injury in a Murine Polymicrobial Sepsis Model ▿"

Article Title: LOX-1 Deletion Improves Neutrophil Responses, Enhances Bacterial Clearance, and Reduces Lung Injury in a Murine Polymicrobial Sepsis Model ▿

Journal: Infection and Immunity

doi: 10.1128/IAI.01317-10

LOX-1 deletion attenuates lung inflammation and injury in sepsis. (A) At 24 h after CLP, lungs from LOX-1 knockout (LOX-1 −/− ) mice and wild-type (WT) mice were collected to examine wet/dry ratios. (B) Lung TNF-α and IL-6 levels
Figure Legend Snippet: LOX-1 deletion attenuates lung inflammation and injury in sepsis. (A) At 24 h after CLP, lungs from LOX-1 knockout (LOX-1 −/− ) mice and wild-type (WT) mice were collected to examine wet/dry ratios. (B) Lung TNF-α and IL-6 levels

Techniques Used: Knock-Out, Mouse Assay

4) Product Images from "Centrally Administered Pertussis Toxin Inhibits Microglia Migration to the Spinal Cord and Prevents Dissemination of Disease in an EAE Mouse Model"

Article Title: Centrally Administered Pertussis Toxin Inhibits Microglia Migration to the Spinal Cord and Prevents Dissemination of Disease in an EAE Mouse Model

Journal: PLoS ONE

doi: 10.1371/journal.pone.0012400

Inflammatory cytokines and cells in the spinal cord of EAE and EAE +PTx icv mice (n = 6/group). IL-17 + /CD4 + cells were detected in the meninges of the spinal cord in the EAE +PTx icv mice (A–C), whereas these cells were diffusely identified in the spinal parenchyma in the EAE mice (D–F). Original magnification ×400. The western blot depicts measures of IL-17 (G), IL-6 (H) and TGF-β (I). In the spinal cord, elevated levels of all three were identified in the EAE mice relative to the EAE +PTx icv mice. Statistical evaluation of optic density (OD) normalized to β-actin was obtained. Mean ± SD are depicted (n = 6 per group). *P
Figure Legend Snippet: Inflammatory cytokines and cells in the spinal cord of EAE and EAE +PTx icv mice (n = 6/group). IL-17 + /CD4 + cells were detected in the meninges of the spinal cord in the EAE +PTx icv mice (A–C), whereas these cells were diffusely identified in the spinal parenchyma in the EAE mice (D–F). Original magnification ×400. The western blot depicts measures of IL-17 (G), IL-6 (H) and TGF-β (I). In the spinal cord, elevated levels of all three were identified in the EAE mice relative to the EAE +PTx icv mice. Statistical evaluation of optic density (OD) normalized to β-actin was obtained. Mean ± SD are depicted (n = 6 per group). *P

Techniques Used: Mouse Assay, Western Blot

5) Product Images from "Nitrate partially inhibits lipopolysaccharide-induced inflammation by maintaining mitochondrial function"

Article Title: Nitrate partially inhibits lipopolysaccharide-induced inflammation by maintaining mitochondrial function

Journal: The Journal of International Medical Research

doi: 10.1177/0300060520902605

Nitrate relieved LPS-induced inflammation. The concentration of IL-6 (a) and TNF-α (b) in the supernatant after 2-hour LPS treatment (n = 4). Western blot of total and phosphorylated levels of IKKα (c) and NF-κB (p65) (d) after 1-hour LPS treatment (n = 3). Data are expressed as mean ± SEM; p
Figure Legend Snippet: Nitrate relieved LPS-induced inflammation. The concentration of IL-6 (a) and TNF-α (b) in the supernatant after 2-hour LPS treatment (n = 4). Western blot of total and phosphorylated levels of IKKα (c) and NF-κB (p65) (d) after 1-hour LPS treatment (n = 3). Data are expressed as mean ± SEM; p

Techniques Used: Concentration Assay, Western Blot

Nitrate did not modulate mitochondrial function or the inflammatory response of macrophages without LPS. After a 2-hour treatment with various concentrations of nitrate (10, 100, and 500 µM), the ΔΨm (a), mtROS (b), and concentrations of IL-6 (c) and TNF-α (d) in the supernatant were detected (n = 3). Data are expressed as mean ± SEM.
Figure Legend Snippet: Nitrate did not modulate mitochondrial function or the inflammatory response of macrophages without LPS. After a 2-hour treatment with various concentrations of nitrate (10, 100, and 500 µM), the ΔΨm (a), mtROS (b), and concentrations of IL-6 (c) and TNF-α (d) in the supernatant were detected (n = 3). Data are expressed as mean ± SEM.

Techniques Used:

6) Product Images from "Autophagy is involved in regulating the immune response of dendritic cells to influenza A (H1N1) pdm09 infection"

Article Title: Autophagy is involved in regulating the immune response of dendritic cells to influenza A (H1N1) pdm09 infection

Journal: Immunology

doi: 10.1111/imm.12587

Autophagy‐deficient bone‐marrow‐derived dendritic cells (BMDCs) exhibited an impaired ability to induce an inflammatory response and an H1N1‐specific T‐cell response in vivo . (a–d) CD11c‐DTR mice received an intraperitoneal injection of diphtheria toxin (16 μg/kg body weight). Twenty‐four hours later, 2 × 10 6 wild‐type (WT) or Beclin‐1 +/− BMDCs were intravenously transferred into these mice, which were then intranasally infected with H1N1 virus (10 2 PFU/mouse) under parenteral anaesthesia. At 2 days post‐infection, the mice were killed, and the levels of interleukin‐6 (IL‐6), tumour necrosis factor‐ α (TNF‐ α ), interferon‐ β (IFN‐ β ) in bronchoalveolar lavage fluid were detected by ELISA (a). At 7 days post‐infection, the mice were killed, and the viral loads in the lungs were visualized by H1N1‐specific immunofluorescent staining (b). Alternatively, the virus infectivity in the lungs was quantified by titration on Madin–Darby canine kidney cells (c), or CD4 + or CD8 + T cells from the spleen were purified and co‐cultured with mitomycin C‐inactivated BMDCs at ratio of 10 : 1, either with or without H1N1 virus at a multiplicity of infection (MOI) of 2 for 24 hr in IL‐2 or IFN‐ γ antibody‐coated plates. Then, the IL‐2‐producing CD4 + T cells or IFN‐ γ ‐producing CD8 + T cells were quantified by ELISpot assay (d). The data are representative of two independent experiments and are expressed as the means ± SEM. Statistical significance was determined using an unpaired Student's t ‐test. * P
Figure Legend Snippet: Autophagy‐deficient bone‐marrow‐derived dendritic cells (BMDCs) exhibited an impaired ability to induce an inflammatory response and an H1N1‐specific T‐cell response in vivo . (a–d) CD11c‐DTR mice received an intraperitoneal injection of diphtheria toxin (16 μg/kg body weight). Twenty‐four hours later, 2 × 10 6 wild‐type (WT) or Beclin‐1 +/− BMDCs were intravenously transferred into these mice, which were then intranasally infected with H1N1 virus (10 2 PFU/mouse) under parenteral anaesthesia. At 2 days post‐infection, the mice were killed, and the levels of interleukin‐6 (IL‐6), tumour necrosis factor‐ α (TNF‐ α ), interferon‐ β (IFN‐ β ) in bronchoalveolar lavage fluid were detected by ELISA (a). At 7 days post‐infection, the mice were killed, and the viral loads in the lungs were visualized by H1N1‐specific immunofluorescent staining (b). Alternatively, the virus infectivity in the lungs was quantified by titration on Madin–Darby canine kidney cells (c), or CD4 + or CD8 + T cells from the spleen were purified and co‐cultured with mitomycin C‐inactivated BMDCs at ratio of 10 : 1, either with or without H1N1 virus at a multiplicity of infection (MOI) of 2 for 24 hr in IL‐2 or IFN‐ γ antibody‐coated plates. Then, the IL‐2‐producing CD4 + T cells or IFN‐ γ ‐producing CD8 + T cells were quantified by ELISpot assay (d). The data are representative of two independent experiments and are expressed as the means ± SEM. Statistical significance was determined using an unpaired Student's t ‐test. * P

Techniques Used: Derivative Assay, In Vivo, Mouse Assay, Injection, Infection, Enzyme-linked Immunosorbent Assay, Staining, Titration, Purification, Cell Culture, Enzyme-linked Immunospot

Beclin‐1 +/− bone‐marrow‐derived dendritic cells (BMDCs) are deficient in maturation and the production of innate and adaptive cytokines upon H1N1virus infection. (a, b) BMDCs from wild‐type (WT) or Beclin‐1 +/− mice were infected with H1N1 virus at a multiplicity of infection (MOI) of 2 for 2 hr. The LC3B‐positive puncta in BMDCs were detected by fluorescence staining (a), and the LC3B‐I/II and p62 proteins were detected by Western blot (b). (c) BMDCs from WT or Beclin‐1 +/− mice were infected with H1N1 virus at an MOI of 2 for 24 hr. The expression of MHC‐II, CD80, CD86 and CD40 was detected by flow cytometry. (d) Statistical analysis of the mean fluorescent intensity (MFI) of (c). (e) BMDCs from WT or Beclin‐1 +/− mice were infected with H1N1 virus at an MOI of 2 for 24 hr. The secretion of interleukin‐6 (IL‐6), tumour necrosis factor‐ α (TNF‐ α ), interferon‐ β (IFN‐ β ), IL‐12p70 and IFN‐ γ was measured by ELISA. (f) The H1N1 virus yield by BMDCs from WT or Beclin‐1 +/− mice at the indicated time‐points after H1N1 virus infection were determined by TCID 50 assay. (g) BMDCs from WT or Beclin‐1 +/− mice were cultured in Hanks' balanced salt solution medium for 2 hr then were infected by H1N1 virus at an MOI of 2 for 24 hr. The secretion of IL‐6, TNF‐ α , IFN‐ β , IL‐12p70 and IFN‐ γ was examined by ELISA. The data are representative of three independent experiments and are expressed as the means ± SEM. Statistical significance was determined using an unpaired Student's t ‐test for (d) and (e), and (g) was analysed by one‐way analysis of variance and Q test. * P
Figure Legend Snippet: Beclin‐1 +/− bone‐marrow‐derived dendritic cells (BMDCs) are deficient in maturation and the production of innate and adaptive cytokines upon H1N1virus infection. (a, b) BMDCs from wild‐type (WT) or Beclin‐1 +/− mice were infected with H1N1 virus at a multiplicity of infection (MOI) of 2 for 2 hr. The LC3B‐positive puncta in BMDCs were detected by fluorescence staining (a), and the LC3B‐I/II and p62 proteins were detected by Western blot (b). (c) BMDCs from WT or Beclin‐1 +/− mice were infected with H1N1 virus at an MOI of 2 for 24 hr. The expression of MHC‐II, CD80, CD86 and CD40 was detected by flow cytometry. (d) Statistical analysis of the mean fluorescent intensity (MFI) of (c). (e) BMDCs from WT or Beclin‐1 +/− mice were infected with H1N1 virus at an MOI of 2 for 24 hr. The secretion of interleukin‐6 (IL‐6), tumour necrosis factor‐ α (TNF‐ α ), interferon‐ β (IFN‐ β ), IL‐12p70 and IFN‐ γ was measured by ELISA. (f) The H1N1 virus yield by BMDCs from WT or Beclin‐1 +/− mice at the indicated time‐points after H1N1 virus infection were determined by TCID 50 assay. (g) BMDCs from WT or Beclin‐1 +/− mice were cultured in Hanks' balanced salt solution medium for 2 hr then were infected by H1N1 virus at an MOI of 2 for 24 hr. The secretion of IL‐6, TNF‐ α , IFN‐ β , IL‐12p70 and IFN‐ γ was examined by ELISA. The data are representative of three independent experiments and are expressed as the means ± SEM. Statistical significance was determined using an unpaired Student's t ‐test for (d) and (e), and (g) was analysed by one‐way analysis of variance and Q test. * P

Techniques Used: Derivative Assay, Infection, Mouse Assay, Fluorescence, Staining, Western Blot, Expressing, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Cell Culture

7) Product Images from "Interleukin 23 Produced by Myeloid Dendritic Cells Contributes to T-Cell Dysfunction in HIV Type 1 Infection by Inducing SOCS1 Expression"

Article Title: Interleukin 23 Produced by Myeloid Dendritic Cells Contributes to T-Cell Dysfunction in HIV Type 1 Infection by Inducing SOCS1 Expression

Journal: The Journal of Infectious Diseases

doi: 10.1093/infdis/jiu523

Human immunodeficiency virus (HIV) gp120 treatment inhibits interleukin 12p70 (IL-12p70) production and increases interleukin 23 (IL-23) production by myeloid dendritic cells (mDCs). A total of 0.5 × 10 6 mDCs were unstimulated (control) or stimulated with 1 µg/mL lipopolysaccharide (LPS) in the presence or absence of gp120 at 1 µg/mL in 50 µL of culture volume. After 24 hours, levels of IL-12p70 ( A ) and interleukin 15 (IL-15; B ) were determined by a enzyme-linked immunosorbent assay (ELISA). A total of 0.1 × 10 6 mDCs were unstimulated (control) or stimulated with 1 µg/mL LPS in the presence or absence of gp120 at 1 µg/mL in 100 µL of culture volume. After 24 hours, levels of interleukin 6 (IL-6; C ) and IL-23 ( D ) were determined by ELISA. All ELISA assays were run in triplicate. Data are mean ± SD (n = 5). Abbreviation: NS, not significant.
Figure Legend Snippet: Human immunodeficiency virus (HIV) gp120 treatment inhibits interleukin 12p70 (IL-12p70) production and increases interleukin 23 (IL-23) production by myeloid dendritic cells (mDCs). A total of 0.5 × 10 6 mDCs were unstimulated (control) or stimulated with 1 µg/mL lipopolysaccharide (LPS) in the presence or absence of gp120 at 1 µg/mL in 50 µL of culture volume. After 24 hours, levels of IL-12p70 ( A ) and interleukin 15 (IL-15; B ) were determined by a enzyme-linked immunosorbent assay (ELISA). A total of 0.1 × 10 6 mDCs were unstimulated (control) or stimulated with 1 µg/mL LPS in the presence or absence of gp120 at 1 µg/mL in 100 µL of culture volume. After 24 hours, levels of interleukin 6 (IL-6; C ) and IL-23 ( D ) were determined by ELISA. All ELISA assays were run in triplicate. Data are mean ± SD (n = 5). Abbreviation: NS, not significant.

Techniques Used: Enzyme-linked Immunosorbent Assay

8) Product Images from "Cyclic mechanical stretch up-regulates hepatoma-derived growth factor expression in cultured rat aortic smooth muscle cells"

Article Title: Cyclic mechanical stretch up-regulates hepatoma-derived growth factor expression in cultured rat aortic smooth muscle cells

Journal: Bioscience Reports

doi: 10.1042/BSR20171398

Effect of HDGF gene-silencing on the constitutive cytokine production in rat aortic smooth muscle cells (SMCs) Cultured SMCs received scramble nucleotides or HDGF siRNA for 24 h, followed by another 24-h incubation. ( A ) Western blotting data confirmed gene-silencing efficiency of siNRA-mediated HDGF gene knockdown after 48-h treatment. Twenty-four hours of conditioned media after HDGF gene knockdown were subjected to ELISA, revealing that HDGF gene modification did not affect TNF-α release ( B ), but significantly reduced IL-6 production ( C ) in SMCs. Data are shown in mean ± SEM; * P
Figure Legend Snippet: Effect of HDGF gene-silencing on the constitutive cytokine production in rat aortic smooth muscle cells (SMCs) Cultured SMCs received scramble nucleotides or HDGF siRNA for 24 h, followed by another 24-h incubation. ( A ) Western blotting data confirmed gene-silencing efficiency of siNRA-mediated HDGF gene knockdown after 48-h treatment. Twenty-four hours of conditioned media after HDGF gene knockdown were subjected to ELISA, revealing that HDGF gene modification did not affect TNF-α release ( B ), but significantly reduced IL-6 production ( C ) in SMCs. Data are shown in mean ± SEM; * P

Techniques Used: Cell Culture, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Modification

Involvement of signaling pathways in cyclic mechanical stretch-increased cytokine production in rat aortic smooth muscle cells (SMCs) SMCs were seeded on fibronectin-coated silicone elastomer chambers till full attachment. The cells were treated with selective kinase inhibitors at 10 μM or 0.1% DMSO solvent control (SC) for 1 h, followed by uniaxial and cyclic 10% stretches at constant frequency (1 Hz) for 24 h. Conditioned media from the stretched SMCs and nonstretching control (NC) were collected after consecutive 24-h stretching and subjected to ELISA detectionof TNF-α ( A ) and IL-6 ( B ). Data are shown in mean ± SEM; * P
Figure Legend Snippet: Involvement of signaling pathways in cyclic mechanical stretch-increased cytokine production in rat aortic smooth muscle cells (SMCs) SMCs were seeded on fibronectin-coated silicone elastomer chambers till full attachment. The cells were treated with selective kinase inhibitors at 10 μM or 0.1% DMSO solvent control (SC) for 1 h, followed by uniaxial and cyclic 10% stretches at constant frequency (1 Hz) for 24 h. Conditioned media from the stretched SMCs and nonstretching control (NC) were collected after consecutive 24-h stretching and subjected to ELISA detectionof TNF-α ( A ) and IL-6 ( B ). Data are shown in mean ± SEM; * P

Techniques Used: Enzyme-linked Immunosorbent Assay

9) Product Images from "Beneficial Effects of Qingzixiaoban Granule on Henoch–Schönlein Purpura Nephritis Mice through Inhibiting Immune Complex Deposition and Th2 Immunodeviation"

Article Title: Beneficial Effects of Qingzixiaoban Granule on Henoch–Schönlein Purpura Nephritis Mice through Inhibiting Immune Complex Deposition and Th2 Immunodeviation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2019/3050248

QZXB GR decreased the levels of CIC, IL-6, renal biochemical parameters in the serum and suppressed immune complex deposition in HSPN mice kidneys. (a) The level of serum CIC in HSPN mice. (b) The level of serum IL-6 in HSPN mice. (c) The level of serum TP in HSPN mice. (d) The level of serum ALB in HSPN mice. (e) The level of serum Cre in HSPN mice. (f) The level of serum BUN in HSPN mice. (g) Representative photomicrographs of IgA deposition (×200). (h) Quantitative scoring of IgA deposition. (i) Representative photomicrographs of IgG deposition (×200). (j) Quantitative scoring of IgG deposition. Data were presented as means ± SEM, n = 8. # P
Figure Legend Snippet: QZXB GR decreased the levels of CIC, IL-6, renal biochemical parameters in the serum and suppressed immune complex deposition in HSPN mice kidneys. (a) The level of serum CIC in HSPN mice. (b) The level of serum IL-6 in HSPN mice. (c) The level of serum TP in HSPN mice. (d) The level of serum ALB in HSPN mice. (e) The level of serum Cre in HSPN mice. (f) The level of serum BUN in HSPN mice. (g) Representative photomicrographs of IgA deposition (×200). (h) Quantitative scoring of IgA deposition. (i) Representative photomicrographs of IgG deposition (×200). (j) Quantitative scoring of IgG deposition. Data were presented as means ± SEM, n = 8. # P

Techniques Used: Mouse Assay

10) Product Images from "The role of the surface on microglia function: implications for central nervous system tissue engineering"

Article Title: The role of the surface on microglia function: implications for central nervous system tissue engineering

Journal: Journal of the Royal Society Interface

doi: 10.1098/rsif.2014.1224

Cytokine release by microglia cultured on P(TMC-CL) substrates. Dot-plot showing the concentration of ( a ) IL-10 and TNFα in cell culture media after 1 or ( b ) 5 days in culture when seeded on P(TMC-CL) surfaces ( n = 5). Panel ( b ) also shows the concentration of IL-6 ( n = 3). ( c ) Release of pro-inflammatory cytokines (TNFα, IL-6) after microglia stimulation with lipopolysacharide (100 ng ml −1 , 3 h, LPS). Asterisks (*) denote statistical significance, p
Figure Legend Snippet: Cytokine release by microglia cultured on P(TMC-CL) substrates. Dot-plot showing the concentration of ( a ) IL-10 and TNFα in cell culture media after 1 or ( b ) 5 days in culture when seeded on P(TMC-CL) surfaces ( n = 5). Panel ( b ) also shows the concentration of IL-6 ( n = 3). ( c ) Release of pro-inflammatory cytokines (TNFα, IL-6) after microglia stimulation with lipopolysacharide (100 ng ml −1 , 3 h, LPS). Asterisks (*) denote statistical significance, p

Techniques Used: Cell Culture, Concentration Assay

11) Product Images from "NMO-IgG and AQP4 Peptide Can Induce Aggravation of EAMG and Immune-Mediated Muscle Weakness"

Article Title: NMO-IgG and AQP4 Peptide Can Induce Aggravation of EAMG and Immune-Mediated Muscle Weakness

Journal: Journal of Immunology Research

doi: 10.1155/2018/5389282

Increased proinflammatory cytokine secretion. Splenocytes derived from EAMG control, EAMG with NMO-Ig, and from naive mice injected with NMO-Ig or AQP4 peptide, or CFA, was activated by anti-CD3. Cell-free supernatants were tested for the cytokine content by specific ELISA. IFN γ was tested after 24 h of stimulation, IL-6 (48 h of stimulation), and IL-10 (after 72 h). Results are the mean of 2 different sets of experiments of 3–5 mice in each group and are expressed as the mean ± SE. ∗ P
Figure Legend Snippet: Increased proinflammatory cytokine secretion. Splenocytes derived from EAMG control, EAMG with NMO-Ig, and from naive mice injected with NMO-Ig or AQP4 peptide, or CFA, was activated by anti-CD3. Cell-free supernatants were tested for the cytokine content by specific ELISA. IFN γ was tested after 24 h of stimulation, IL-6 (48 h of stimulation), and IL-10 (after 72 h). Results are the mean of 2 different sets of experiments of 3–5 mice in each group and are expressed as the mean ± SE. ∗ P

Techniques Used: Derivative Assay, Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay

12) Product Images from "Endocytosis of Albumin by Podocytes Elicits an Inflammatory Response and Induces Apoptotic Cell Death"

Article Title: Endocytosis of Albumin by Podocytes Elicits an Inflammatory Response and Induces Apoptotic Cell Death

Journal: PLoS ONE

doi: 10.1371/journal.pone.0054817

Albumin exposure modulates pro-inflammatory cytokine expression and release in human podocytes. A , Time course of IL-1β RNA levels in podocytes treated with 5 mg/ml recombinant low endotoxin human albumin (closed bars) or 5 mg/ml dextran (open bars). * denotes P = 0.0002 compared to dextran controls. B , Time course of TNF RNA expression in podocytes treated with 5 mg/ml human albumin (closed bars) or 5 mg/ml dextran (open bars). * denotes P = 0.01 compared to dextran control. C , IL-6 RNA levels in podocytes treated with 5 mg/ml human albumin (closed bars) or 5 mg/ml dextran (open bars). * denotes P = 0.008 compared to dextran control. D, Amount of IL-1β normalized to total cellular protein released into the medium after treatment of podocytes with 5 mg/ml recombinant human albumin (closed bars) or 5 mg/ml dextran (open bars) for varying amounts of time. * denotes P = 0.005 compared to dextran treated controls. E , Amount of TNF normalized to total cellular protein released into the medium by podocytes after treatment with albumin (closed bars) or dextran (open bars) for varying amounts of time. * denotes P = 0.003 compared to dextran treated control cells. F , Levels of IL-6 normalized to total cellular protein released into the medium by podocytes treated with albumin (closed bars) or dextran (open bars) for varying amounts of time. * denotes P
Figure Legend Snippet: Albumin exposure modulates pro-inflammatory cytokine expression and release in human podocytes. A , Time course of IL-1β RNA levels in podocytes treated with 5 mg/ml recombinant low endotoxin human albumin (closed bars) or 5 mg/ml dextran (open bars). * denotes P = 0.0002 compared to dextran controls. B , Time course of TNF RNA expression in podocytes treated with 5 mg/ml human albumin (closed bars) or 5 mg/ml dextran (open bars). * denotes P = 0.01 compared to dextran control. C , IL-6 RNA levels in podocytes treated with 5 mg/ml human albumin (closed bars) or 5 mg/ml dextran (open bars). * denotes P = 0.008 compared to dextran control. D, Amount of IL-1β normalized to total cellular protein released into the medium after treatment of podocytes with 5 mg/ml recombinant human albumin (closed bars) or 5 mg/ml dextran (open bars) for varying amounts of time. * denotes P = 0.005 compared to dextran treated controls. E , Amount of TNF normalized to total cellular protein released into the medium by podocytes after treatment with albumin (closed bars) or dextran (open bars) for varying amounts of time. * denotes P = 0.003 compared to dextran treated control cells. F , Levels of IL-6 normalized to total cellular protein released into the medium by podocytes treated with albumin (closed bars) or dextran (open bars) for varying amounts of time. * denotes P

Techniques Used: Expressing, Recombinant, RNA Expression

Albumin overload induces proteinuria and upregulates pro-inflammatory cytokine expression in isolated mouse glomeruli. A , Urinary protein normalized to urinary creatinine in mice injected with saline (open bar) or low endotoxin bovine serum albumin (BSA; closed bar; n = 4 animals per group). * denotes P = 0.004 compared to saline injected controls. B , IL-1β expression in isolated glomeruli from mice injected with saline (open bar) or BSA (closed bar; n = 4 animals per group). * denotes P = 0.02 compared to controls. C , TNF expression in isolated glomeruli from mice injected with saline (open bar) or BSA (closed bar; n = 4 animals per group). * denotes P = 0.02 compared to controls. D , IL-6 expression in isolated glomeruli from mice injected with saline (open bar) or BSA (closed bar; n = 4 animals per group).
Figure Legend Snippet: Albumin overload induces proteinuria and upregulates pro-inflammatory cytokine expression in isolated mouse glomeruli. A , Urinary protein normalized to urinary creatinine in mice injected with saline (open bar) or low endotoxin bovine serum albumin (BSA; closed bar; n = 4 animals per group). * denotes P = 0.004 compared to saline injected controls. B , IL-1β expression in isolated glomeruli from mice injected with saline (open bar) or BSA (closed bar; n = 4 animals per group). * denotes P = 0.02 compared to controls. C , TNF expression in isolated glomeruli from mice injected with saline (open bar) or BSA (closed bar; n = 4 animals per group). * denotes P = 0.02 compared to controls. D , IL-6 expression in isolated glomeruli from mice injected with saline (open bar) or BSA (closed bar; n = 4 animals per group).

Techniques Used: Expressing, Isolation, Mouse Assay, Injection

13) Product Images from "Therapeutic effects of pegylated-interferon-α2a in a mouse model of multiple sclerosis"

Article Title: Therapeutic effects of pegylated-interferon-α2a in a mouse model of multiple sclerosis

Journal: Central-European Journal of Immunology

doi: 10.5114/ceji.2018.74868

The effect of Peg-IFN α-2a on serum IL-6 concentration. It was observed that treatment with Peg-IFN α-2a significantly decreased IL-6 concentration compared to the control group ( p = 0.046)
Figure Legend Snippet: The effect of Peg-IFN α-2a on serum IL-6 concentration. It was observed that treatment with Peg-IFN α-2a significantly decreased IL-6 concentration compared to the control group ( p = 0.046)

Techniques Used: Concentration Assay

14) Product Images from "Anti-Inflammatory and Cytostatic Activities of a Parthenolide-Like Sesquiterpene Lactone from Cota palaestina subsp. syriaca"

Article Title: Anti-Inflammatory and Cytostatic Activities of a Parthenolide-Like Sesquiterpene Lactone from Cota palaestina subsp. syriaca

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2015/474597

Identification of the anti-inflammatory component “K100” in Cota palaestina : (a) effect of II.1, II.2, II.3, and II.4 fractions (10 μ g/mL). (b) Effect of subfractions (II.2.2, II.2.3, II.2.5, and II.2.7) at 10 μ g/mL on IL-6 production and (c) dose-dependent inhibition of IL-6 production by fraction II.2.5 at 1, 3, 6, 9, and 12 μ g/mL. (d) Purification and structure of the anti-inflammatory component “K100” in Cota palaestina . Diagram summarizing the fractionation of Cota palaestina extract. (−) indicates no suppression of IL-6 production by ET-treated SCp2 cells and (+) indicates suppression of IL-6 production. Chemical structure of K100, the germacranolide 1- β ,10- α -Epoxy-6-hydroxy-1,10H-inunolide. Statistical significance is represented by ( ∗∗∗ ) asterisk indicating significant difference at p
Figure Legend Snippet: Identification of the anti-inflammatory component “K100” in Cota palaestina : (a) effect of II.1, II.2, II.3, and II.4 fractions (10 μ g/mL). (b) Effect of subfractions (II.2.2, II.2.3, II.2.5, and II.2.7) at 10 μ g/mL on IL-6 production and (c) dose-dependent inhibition of IL-6 production by fraction II.2.5 at 1, 3, 6, 9, and 12 μ g/mL. (d) Purification and structure of the anti-inflammatory component “K100” in Cota palaestina . Diagram summarizing the fractionation of Cota palaestina extract. (−) indicates no suppression of IL-6 production by ET-treated SCp2 cells and (+) indicates suppression of IL-6 production. Chemical structure of K100, the germacranolide 1- β ,10- α -Epoxy-6-hydroxy-1,10H-inunolide. Statistical significance is represented by ( ∗∗∗ ) asterisk indicating significant difference at p

Techniques Used: Inhibition, Purification, Fractionation

K100 inhibits IL-6, NO, and MMP-9 production in ET-treated SCp2 cells. Cells were treated with 0, 5, 10, 15, and 20 μ M of K100 and media samples were collected 24 hrs after ET-stimulation and analyzed for their (a) IL-6 secretion, (b) NO production, and (c) MMP-9 and MMP-2 production. Zymograms were analyzed by Gel documentation (Bio-Rad) using the software Quantity 1. Statistical significance is represented by ( ∗∗∗ ) asterisk indicating significant difference at p
Figure Legend Snippet: K100 inhibits IL-6, NO, and MMP-9 production in ET-treated SCp2 cells. Cells were treated with 0, 5, 10, 15, and 20 μ M of K100 and media samples were collected 24 hrs after ET-stimulation and analyzed for their (a) IL-6 secretion, (b) NO production, and (c) MMP-9 and MMP-2 production. Zymograms were analyzed by Gel documentation (Bio-Rad) using the software Quantity 1. Statistical significance is represented by ( ∗∗∗ ) asterisk indicating significant difference at p

Techniques Used: Software

15) Product Images from "Role of microRNAs in Resveratrol-Mediated Mitigation of Colitis-Associated Tumorigenesis in ApcMin/+"

Article Title: Role of microRNAs in Resveratrol-Mediated Mitigation of Colitis-Associated Tumorigenesis in ApcMin/+

Journal: The Journal of Pharmacology and Experimental Therapeutics

doi: 10.1124/jpet.114.213306

Resveratrol reduces the concentration of proinflammatory cytokines in the small intestine. The effects of resveratrol compared with vehicle on the concentration of IL-6 (A) and TNF- α (B) in pooled sections 2 and 3 of the small intestine of DSS-exposed
Figure Legend Snippet: Resveratrol reduces the concentration of proinflammatory cytokines in the small intestine. The effects of resveratrol compared with vehicle on the concentration of IL-6 (A) and TNF- α (B) in pooled sections 2 and 3 of the small intestine of DSS-exposed

Techniques Used: Concentration Assay

16) Product Images from "Role of gp91phox in hepatic macrophage programming and alcoholic liver disease"

Article Title: Role of gp91phox in hepatic macrophage programming and alcoholic liver disease

Journal: Hepatology Communications

doi: 10.1002/hep4.1078

gp91 phox−/− mice exhibit an increased inflammatory response to chronic alcohol treatment. Female WT and gp91 phox−/− mice were treated as described in Fig. 1 . (A) Serum levels of TNF‐α, IFN‐γ, and IL‐6 were determined by enzyme‐linked immunosorbent assay. * P
Figure Legend Snippet: gp91 phox−/− mice exhibit an increased inflammatory response to chronic alcohol treatment. Female WT and gp91 phox−/− mice were treated as described in Fig. 1 . (A) Serum levels of TNF‐α, IFN‐γ, and IL‐6 were determined by enzyme‐linked immunosorbent assay. * P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

17) Product Images from "NMO-IgG and AQP4 Peptide Can Induce Aggravation of EAMG and Immune-Mediated Muscle Weakness"

Article Title: NMO-IgG and AQP4 Peptide Can Induce Aggravation of EAMG and Immune-Mediated Muscle Weakness

Journal: Journal of Immunology Research

doi: 10.1155/2018/5389282

Increased proinflammatory cytokine secretion. Splenocytes derived from EAMG control, EAMG with NMO-Ig, and from naive mice injected with NMO-Ig or AQP4 peptide, or CFA, was activated by anti-CD3. Cell-free supernatants were tested for the cytokine content by specific ELISA. IFN γ was tested after 24 h of stimulation, IL-6 (48 h of stimulation), and IL-10 (after 72 h). Results are the mean of 2 different sets of experiments of 3–5 mice in each group and are expressed as the mean ± SE. ∗ P
Figure Legend Snippet: Increased proinflammatory cytokine secretion. Splenocytes derived from EAMG control, EAMG with NMO-Ig, and from naive mice injected with NMO-Ig or AQP4 peptide, or CFA, was activated by anti-CD3. Cell-free supernatants were tested for the cytokine content by specific ELISA. IFN γ was tested after 24 h of stimulation, IL-6 (48 h of stimulation), and IL-10 (after 72 h). Results are the mean of 2 different sets of experiments of 3–5 mice in each group and are expressed as the mean ± SE. ∗ P

Techniques Used: Derivative Assay, Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay

18) Product Images from "Cyclic mechanical stretch up-regulates hepatoma-derived growth factor expression in cultured rat aortic smooth muscle cells"

Article Title: Cyclic mechanical stretch up-regulates hepatoma-derived growth factor expression in cultured rat aortic smooth muscle cells

Journal: Bioscience Reports

doi: 10.1042/BSR20171398

Effect of HDGF gene-silencing on the constitutive cytokine production in rat aortic smooth muscle cells (SMCs) Cultured SMCs received scramble nucleotides or HDGF siRNA for 24 h, followed by another 24-h incubation. ( A ) Western blotting data confirmed gene-silencing efficiency of siNRA-mediated HDGF gene knockdown after 48-h treatment. Twenty-four hours of conditioned media after HDGF gene knockdown were subjected to ELISA, revealing that HDGF gene modification did not affect TNF-α release ( B ), but significantly reduced IL-6 production ( C ) in SMCs. Data are shown in mean ± SEM; * P
Figure Legend Snippet: Effect of HDGF gene-silencing on the constitutive cytokine production in rat aortic smooth muscle cells (SMCs) Cultured SMCs received scramble nucleotides or HDGF siRNA for 24 h, followed by another 24-h incubation. ( A ) Western blotting data confirmed gene-silencing efficiency of siNRA-mediated HDGF gene knockdown after 48-h treatment. Twenty-four hours of conditioned media after HDGF gene knockdown were subjected to ELISA, revealing that HDGF gene modification did not affect TNF-α release ( B ), but significantly reduced IL-6 production ( C ) in SMCs. Data are shown in mean ± SEM; * P

Techniques Used: Cell Culture, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Modification

Involvement of signaling pathways in cyclic mechanical stretch-increased cytokine production in rat aortic smooth muscle cells (SMCs) SMCs were seeded on fibronectin-coated silicone elastomer chambers till full attachment. The cells were treated with selective kinase inhibitors at 10 μM or 0.1% DMSO solvent control (SC) for 1 h, followed by uniaxial and cyclic 10% stretches at constant frequency (1 Hz) for 24 h. Conditioned media from the stretched SMCs and nonstretching control (NC) were collected after consecutive 24-h stretching and subjected to ELISA detectionof TNF-α ( A ) and IL-6 ( B ). Data are shown in mean ± SEM; * P
Figure Legend Snippet: Involvement of signaling pathways in cyclic mechanical stretch-increased cytokine production in rat aortic smooth muscle cells (SMCs) SMCs were seeded on fibronectin-coated silicone elastomer chambers till full attachment. The cells were treated with selective kinase inhibitors at 10 μM or 0.1% DMSO solvent control (SC) for 1 h, followed by uniaxial and cyclic 10% stretches at constant frequency (1 Hz) for 24 h. Conditioned media from the stretched SMCs and nonstretching control (NC) were collected after consecutive 24-h stretching and subjected to ELISA detectionof TNF-α ( A ) and IL-6 ( B ). Data are shown in mean ± SEM; * P

Techniques Used: Enzyme-linked Immunosorbent Assay

19) Product Images from "Conjugated bilirubin affects cytokine profiles in hepatitis A virus infection by modulating function of signal transducer and activator of transcription factors"

Article Title: Conjugated bilirubin affects cytokine profiles in hepatitis A virus infection by modulating function of signal transducer and activator of transcription factors

Journal: Immunology

doi: 10.1111/imm.12336

CB levels were differentially associated with IL-8 and IL-6 secretion during HAV infection
Figure Legend Snippet: CB levels were differentially associated with IL-8 and IL-6 secretion during HAV infection

Techniques Used: Infection

Interleukin-8 (IL-8) and IL-6 were differentially regulated by conjugated bilirubin in different hepatitis A virus (HAV) -induced clinical courses. ELISAs were performed to determine the concentrations of cytokines in serum samples from patients with
Figure Legend Snippet: Interleukin-8 (IL-8) and IL-6 were differentially regulated by conjugated bilirubin in different hepatitis A virus (HAV) -induced clinical courses. ELISAs were performed to determine the concentrations of cytokines in serum samples from patients with

Techniques Used:

High concentration of conjugated bilirubin (CB) resulted in interleukin-6 (IL-6) and tumour necrosis factor- α (TNF- α ) secretion in vitro in lymphoid cells from hepatitis A virus (HAV) -infected patients. Peripheral blood lymphoid cells
Figure Legend Snippet: High concentration of conjugated bilirubin (CB) resulted in interleukin-6 (IL-6) and tumour necrosis factor- α (TNF- α ) secretion in vitro in lymphoid cells from hepatitis A virus (HAV) -infected patients. Peripheral blood lymphoid cells

Techniques Used: Concentration Assay, In Vitro, Infection

20) Product Images from "Metabolic Syndrome Exacerbates Inflammation and Bone Loss in Periodontitis"

Article Title: Metabolic Syndrome Exacerbates Inflammation and Bone Loss in Periodontitis

Journal: Journal of Dental Research

doi: 10.1177/0022034514561658

Palmitic acid (PA) enhances lipopolysaccharide (LPS)–stimulated interleukin 6 (IL-6) secretion from bone marrow–derived macrophages, which were treated with different concentrations of LPS, 100 µM of PA, or both LPS and PA for 24 h. After the treatment, IL-6 in culture medium was quantified via ELISA. The data are presented as means ± SD of 1 of 3 experiments with similar results.
Figure Legend Snippet: Palmitic acid (PA) enhances lipopolysaccharide (LPS)–stimulated interleukin 6 (IL-6) secretion from bone marrow–derived macrophages, which were treated with different concentrations of LPS, 100 µM of PA, or both LPS and PA for 24 h. After the treatment, IL-6 in culture medium was quantified via ELISA. The data are presented as means ± SD of 1 of 3 experiments with similar results.

Techniques Used: Derivative Assay, Enzyme-linked Immunosorbent Assay

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Enzyme-linked Immunosorbent Assay:

Article Title: Centrally Administered Pertussis Toxin Inhibits Microglia Migration to the Spinal Cord and Prevents Dissemination of Disease in an EAE Mouse Model
Article Snippet: .. Supernatants were collected and aliquoted in 96-well plate precoated with antibodies to Interferon γ (IFN- γ), Tumor Necrosis Factor α (TNF-α), Interleukin-2 (IL-2), Interleukin-4 (IL-4), Interleukin-6 (IL-6) and Interleukin-10 (IL-10) (ELISA Max™ Set Deluxe, BioLegend Inc. San Diego, CA). .. Optical density was measured at 450 nm on Model 680 Microplate Reader (Bio-Rad Laboratories, Corston,UK).

Article Title: Cyclic mechanical stretch up-regulates hepatoma-derived growth factor expression in cultured rat aortic smooth muscle cells
Article Snippet: .. The soluble cytokines including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were measured by using commercially available ELISA detection kits (Biolegend, San Diego, CA) according to the manufacturer’s instructions. .. RNA interference of HDGF To determine the role of HDGF in constitutive production of cytokines in SMCs, cells were transfected with either small interfering RNA (siRNA) against HDGF gene or scramble RNA control at 100 nM (Ambion/Invitrogen, Carlsbad, CA) by Lipofectamine 2000 (Invitrogen, Grand Island, NY).

Article Title: LOX-1 Deletion Improves Neutrophil Responses, Enhances Bacterial Clearance, and Reduces Lung Injury in a Murine Polymicrobial Sepsis Model ▿
Article Snippet: .. Concentrations of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in plasma or lung homogenates were determined using enzyme-linked immunosorbent assay (ELISA) kits (Biolegend). .. In vitro neutrophil phagocytosis assays were performed as described previously ( , ) using a Vybrant phagocytosis assay kit (Invitrogen).

Article Title: Autophagy is involved in regulating the immune response of dendritic cells to influenza A (H1N1) pdm09 infection
Article Snippet: .. The levels of interleukin‐6 (IL‐6), tumour necrosis factor‐ Biotechnology (TNF‐ α ), IFN‐ β , IL‐12p70 and IFN‐ γ in the culture supernatant (all from Biolegend, San Diego, CA) were measured by ELISA. .. Wild‐type BMDCs were infected with H1N1 viruses at an MOI of 2 with or without 100 n m bafilomycin A1 for 24 hr.

Article Title: Cathelicidin Antimicrobial Peptide Expression Is Not Induced or Required for Bacterial Clearance during Salmonella enterica Infection of Human Monocyte-Derived Macrophages
Article Snippet: .. The proinflammatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) were detected in cell culture supernatant by enzyme-linked immunosorbent assay (ELISA) (Biolegend, San Diego, CA, and R & D Systems, Minneapolis, MN, respectively). .. For LPS stimulation experiments, MDMs were incubated in medium containing 100 ng/ml purified S .

Article Title: A Low-Cost Mechanical Stretching Device for Uniaxial Strain of Cells: A Platform for Pedagogy in Mechanobiology
Article Snippet: .. Following 4 h of incubation, the media was replaced with either regular or 0.3 ng/mL interferon-gamma (IFN-γ) and 0.3 ng/mL lipopolysaccharide (LPS) containing media then cyclically stretched at a 10% stretch amplitude for a period of 18 h. Following stretch, supernatants were collected and analyzed for the presence of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) cytokine secretion using ELISA kits (BioLegend) following the manufacturer's instructions. .. Neonatal rat ventricular myocytes (NRVM) were harvested from two-day-old neonatal Sprague-Dawley rats (Charles River Laboratories, Wilmington, MA), as previously described [ ].

Article Title: Interleukin 23 Produced by Myeloid Dendritic Cells Contributes to T-Cell Dysfunction in HIV Type 1 Infection by Inducing SOCS1 Expression
Article Snippet: .. In some experiments, DCs were stimulated with LPS (1 µg/mL) for 24 hours in the presence or absence of gp120 (1 µg/mL), and concentrations of IL12-p70, interleukin 6 (IL-6), interleukin 15 (IL-15), and IL-23 in culture supernatants were measured by ELISA (Biolegend). .. Expression of CD80, CD83, and HLA-DR was measured by incubating mDCs with allophycocyanin (APC)–anti-CD83, phycoerythrin–anti-HLA-DR, and AF488–anti-CD80 (all Biolegend) on ice for 30 minutes.

Incubation:

Article Title: A Low-Cost Mechanical Stretching Device for Uniaxial Strain of Cells: A Platform for Pedagogy in Mechanobiology
Article Snippet: .. Following 4 h of incubation, the media was replaced with either regular or 0.3 ng/mL interferon-gamma (IFN-γ) and 0.3 ng/mL lipopolysaccharide (LPS) containing media then cyclically stretched at a 10% stretch amplitude for a period of 18 h. Following stretch, supernatants were collected and analyzed for the presence of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) cytokine secretion using ELISA kits (BioLegend) following the manufacturer's instructions. .. Neonatal rat ventricular myocytes (NRVM) were harvested from two-day-old neonatal Sprague-Dawley rats (Charles River Laboratories, Wilmington, MA), as previously described [ ].

Cell Culture:

Article Title: Cathelicidin Antimicrobial Peptide Expression Is Not Induced or Required for Bacterial Clearance during Salmonella enterica Infection of Human Monocyte-Derived Macrophages
Article Snippet: .. The proinflammatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) were detected in cell culture supernatant by enzyme-linked immunosorbent assay (ELISA) (Biolegend, San Diego, CA, and R & D Systems, Minneapolis, MN, respectively). .. For LPS stimulation experiments, MDMs were incubated in medium containing 100 ng/ml purified S .

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    BioLegend il 1β
    The effect of farrerol on pro-inflammatory mediator’s production in LPS-stimulated mMECs. MMECs were divided into six groups: NT group, Farrerol group, LPS group, LPS + farrerol (90, 110, 130 μM) group. The mRNA levels of TNF-α ( A ); IL-6 ( B ) and <t>IL-1β</t> ( C ) were measured by real-time PCR. The protein levels of iNOS ( D , E ) and COX-2 ( D , F ) were measured by Western blot. Data are presented as mean ± SEM ( n = 3). # p
    Il 1β, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 419 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend interleukin 6
    Levels of proinflammatory cytokines released from S . Typhi- or S . Typhimurium-infected MDMs. Shown are levels of TNF-α (A) and <t>IL-6</t> (B) released from MDMs after 2.5 and 24 h of S . Typhi infection and TNF-α (C) and IL-6 (D) released from MDMs after 2.5 and 24 h of S . Typhimurium infection. Statistical significance was determined by Student's t test comparing cytokine levels between LPS modification mutant-infected samples and the WT-infected sample collected at that time point. *, P
    Interleukin 6, supplied by BioLegend, used in various techniques. Bioz Stars score: 93/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    BioLegend il 12p40
    DNA-PKcs is involved in the IL-6 and IL-12 response to CpG-ODN. Bone marrow-derived dendritic cells (DCs) from WT, DNA-PKcs −/− , TLR9 −/− and DNA-PKcs −/−/ TLR9 −/− mice were harvested based on their natural status: adhesion and suspension. ( A ). Combined DCs (adhesion and suspension) at day 7.5 were subjected to flow cytometry. The levels of CD11b, CD11c and MHC-II expression on DCs were determined. ( B–M ). Adhesion or suspension DCs at day 6.5 were seeded in 96-well plates at 1×10 5 /well in triplicate, and then treated with CpG-ODN (CpG, 0.2, 0.73, 2.2 or 7.3 µg/ml), LPS (0.1 µg/ml), R848 (0.125 µg/ml), or left untreated for indicated time durations. The levels of IL-6 ( B–G ) and <t>IL-12p40</t> ( H–M ) were determined by ELISA using IL-6 or IL-12p40 ELISA kits. Similar results were obtained in at least 3 independent experiments. Note: bars represent the average of triplicates ± SD. * p
    Il 12p40, supplied by BioLegend, used in various techniques. Bioz Stars score: 93/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The effect of farrerol on pro-inflammatory mediator’s production in LPS-stimulated mMECs. MMECs were divided into six groups: NT group, Farrerol group, LPS group, LPS + farrerol (90, 110, 130 μM) group. The mRNA levels of TNF-α ( A ); IL-6 ( B ) and IL-1β ( C ) were measured by real-time PCR. The protein levels of iNOS ( D , E ) and COX-2 ( D , F ) were measured by Western blot. Data are presented as mean ± SEM ( n = 3). # p

    Journal: International Journal of Molecular Sciences

    Article Title: Farrerol Relieve Lipopolysaccharide (LPS)-Induced Mastitis by Inhibiting AKT/NF-κB p65, ERK1/2 and P38 Signaling Pathway

    doi: 10.3390/ijms19061770

    Figure Lengend Snippet: The effect of farrerol on pro-inflammatory mediator’s production in LPS-stimulated mMECs. MMECs were divided into six groups: NT group, Farrerol group, LPS group, LPS + farrerol (90, 110, 130 μM) group. The mRNA levels of TNF-α ( A ); IL-6 ( B ) and IL-1β ( C ) were measured by real-time PCR. The protein levels of iNOS ( D , E ) and COX-2 ( D , F ) were measured by Western blot. Data are presented as mean ± SEM ( n = 3). # p

    Article Snippet: The levels of TNF-α, IL-6 and IL-1β in mammary glands were determined using the mouse ELISA kits (Biolegend, San Diego, CA, USA) according to the manufacturer’s instructions.

    Techniques: Real-time Polymerase Chain Reaction, Western Blot

    The levels of TNF-α ( A ); IL-6 ( B ) and IL-1β ( C ) in the homogenate of mouse mammary glands including NT group, LPS group and LPS + farrerol (20, 30, 40 mg/kg) group; The protein levels of iNOS ( D , E ) and COX-2 ( D , F ) were measured by Western blot. Data are presented as mean ± SEM ( n = 3). # p

    Journal: International Journal of Molecular Sciences

    Article Title: Farrerol Relieve Lipopolysaccharide (LPS)-Induced Mastitis by Inhibiting AKT/NF-κB p65, ERK1/2 and P38 Signaling Pathway

    doi: 10.3390/ijms19061770

    Figure Lengend Snippet: The levels of TNF-α ( A ); IL-6 ( B ) and IL-1β ( C ) in the homogenate of mouse mammary glands including NT group, LPS group and LPS + farrerol (20, 30, 40 mg/kg) group; The protein levels of iNOS ( D , E ) and COX-2 ( D , F ) were measured by Western blot. Data are presented as mean ± SEM ( n = 3). # p

    Article Snippet: The levels of TNF-α, IL-6 and IL-1β in mammary glands were determined using the mouse ELISA kits (Biolegend, San Diego, CA, USA) according to the manufacturer’s instructions.

    Techniques: Western Blot

    Infection of monocyte-derived macrophages (MDMs) and unactivated CD4 + T-cells with T/F viruses reproduce the results obtained with the R5-tropic (BAL) and the X4-tropic (NL4-3) laboratory strains. (A) Expression of IL-1β, IL-8, IL-6, and IL-10 in supernatants from uninfected (Un) and R5-tropic T/F-infected (In) MDMs. Data represent mean ± SD from three independent donors. Data were analyzed by two-way ANOVA followed by Tukey’s post-test. * p

    Journal: Frontiers in Immunology

    Article Title: Interaction Between Macrophage Migration Inhibitory Factor and CD74 in Human Immunodeficiency Virus Type I Infected Primary Monocyte-Derived Macrophages Triggers the Production of Proinflammatory Mediators and Enhances Infection of Unactivated CD4+ T Cells

    doi: 10.3389/fimmu.2018.01494

    Figure Lengend Snippet: Infection of monocyte-derived macrophages (MDMs) and unactivated CD4 + T-cells with T/F viruses reproduce the results obtained with the R5-tropic (BAL) and the X4-tropic (NL4-3) laboratory strains. (A) Expression of IL-1β, IL-8, IL-6, and IL-10 in supernatants from uninfected (Un) and R5-tropic T/F-infected (In) MDMs. Data represent mean ± SD from three independent donors. Data were analyzed by two-way ANOVA followed by Tukey’s post-test. * p

    Article Snippet: Cytokine Quantitation The levels of the following cytokines were evaluated in MDM supernatants using commercially available ELISA sets: IL-8, IL-6, IL-1β, TNFα, IL-10 (ELISA MAX Deluxe kits, BioLegend) and sICAM (DouSet ELISA, R & D Systems).

    Techniques: Infection, Derivative Assay, Expressing

    Identification of cytokines as responsible for enhancing human immunodeficiency virus type I (HIV-1) infection in unactivated CD4 + T-cells. (A) Unactivated CD4 + T-cells were stimulated with different combinations of cytokines for 72 h. Then, cells were infected and p24 antigen production was evaluated at days 4 and 7 post-infection. Each condition was compared with the corresponding RPMI condition (negative control). As a positive control, PHA stimulation was used. Percentage of living CD4 + T-cells (B) and percentage of infected (GFP + ) CD4 + T-cells (C) after stimulation with the denoted treatments are shown. Data represent mean ± SD from four independent donors evaluated in duplicate. Concentrations of cytokines used in these experiments corresponded to the average concentrations found in monocyte-derived macrophage (MDM) supernatants stimulated with 25 ng/ml macrophage migration inhibitory factor (MIF) (peak effect) as follows: 250 pg/ml IL-6, 9,000 pg/ml IL-8, 1,400 pg/ml TNF-α, and 20 pg/ml IL-1β. (D) Neutralization of IL-8, IL-6, IL-1 β, and TNFα biological activity with monoclonal neutralizing antibodies. Primary CD4 + T-cells were incubated with supernatants derived from the 25 ng/ml MIF-treated HIV-infected MDM neutralized previously with 18 µg/ml anti-IL-8, 20 ng/ml anti-IL-6, 2 µg/ml anti-IL-1β, and 2 µg/ml anti-TNFα antibodies. Non-neutralized and isotype control antibody conditions were tested for comparison. Also, RPMI and PHA controls were included. Viral production was evaluated at day 4 post-infection. Data were analyzed by one-way ANOVA followed by Dunnett’s post-test (all conditions versus the corresponding RMPI control) in (A) and Tukey’s post-test in (D) . * p

    Journal: Frontiers in Immunology

    Article Title: Interaction Between Macrophage Migration Inhibitory Factor and CD74 in Human Immunodeficiency Virus Type I Infected Primary Monocyte-Derived Macrophages Triggers the Production of Proinflammatory Mediators and Enhances Infection of Unactivated CD4+ T Cells

    doi: 10.3389/fimmu.2018.01494

    Figure Lengend Snippet: Identification of cytokines as responsible for enhancing human immunodeficiency virus type I (HIV-1) infection in unactivated CD4 + T-cells. (A) Unactivated CD4 + T-cells were stimulated with different combinations of cytokines for 72 h. Then, cells were infected and p24 antigen production was evaluated at days 4 and 7 post-infection. Each condition was compared with the corresponding RPMI condition (negative control). As a positive control, PHA stimulation was used. Percentage of living CD4 + T-cells (B) and percentage of infected (GFP + ) CD4 + T-cells (C) after stimulation with the denoted treatments are shown. Data represent mean ± SD from four independent donors evaluated in duplicate. Concentrations of cytokines used in these experiments corresponded to the average concentrations found in monocyte-derived macrophage (MDM) supernatants stimulated with 25 ng/ml macrophage migration inhibitory factor (MIF) (peak effect) as follows: 250 pg/ml IL-6, 9,000 pg/ml IL-8, 1,400 pg/ml TNF-α, and 20 pg/ml IL-1β. (D) Neutralization of IL-8, IL-6, IL-1 β, and TNFα biological activity with monoclonal neutralizing antibodies. Primary CD4 + T-cells were incubated with supernatants derived from the 25 ng/ml MIF-treated HIV-infected MDM neutralized previously with 18 µg/ml anti-IL-8, 20 ng/ml anti-IL-6, 2 µg/ml anti-IL-1β, and 2 µg/ml anti-TNFα antibodies. Non-neutralized and isotype control antibody conditions were tested for comparison. Also, RPMI and PHA controls were included. Viral production was evaluated at day 4 post-infection. Data were analyzed by one-way ANOVA followed by Dunnett’s post-test (all conditions versus the corresponding RMPI control) in (A) and Tukey’s post-test in (D) . * p

    Article Snippet: Cytokine Quantitation The levels of the following cytokines were evaluated in MDM supernatants using commercially available ELISA sets: IL-8, IL-6, IL-1β, TNFα, IL-10 (ELISA MAX Deluxe kits, BioLegend) and sICAM (DouSet ELISA, R & D Systems).

    Techniques: Infection, Negative Control, Positive Control, Derivative Assay, Migration, Neutralization, Activity Assay, Incubation

    Expression of cytokines after macrophage migration inhibitory factor (MIF) stimulation in primary human immunodeficiency virus (HIV)-infected and uninfected monocyte-derived macrophages (MDMs). (A) Expression of IL-8, IL-6, IL-1β, TNF-α, sICAM, and IL-10 in supernatants from HIV-infected (In) and uninfected (Un) MDMs obtained from one representative healthy donor. (B) Data combined from six independent experiments (donors), each evaluated in triplicate. Here, data are shown as the ratio between cytokine concentrations found under the infection condition versus the uninfected counterpart. Cells were stimulated with MIF as follows: 0, 1, 10, or 25 ng/ml. Data shown in the gray boxes depict CD74 blocking (10 ng/ml of αCD74 or the corresponding isotype control) followed by MIF stimulation (1 or 25 ng/ml as denoted). Data represent the mean ± SD. Data were analyzed by one-way ANOVA followed by Tukey’s post-test. * p

    Journal: Frontiers in Immunology

    Article Title: Interaction Between Macrophage Migration Inhibitory Factor and CD74 in Human Immunodeficiency Virus Type I Infected Primary Monocyte-Derived Macrophages Triggers the Production of Proinflammatory Mediators and Enhances Infection of Unactivated CD4+ T Cells

    doi: 10.3389/fimmu.2018.01494

    Figure Lengend Snippet: Expression of cytokines after macrophage migration inhibitory factor (MIF) stimulation in primary human immunodeficiency virus (HIV)-infected and uninfected monocyte-derived macrophages (MDMs). (A) Expression of IL-8, IL-6, IL-1β, TNF-α, sICAM, and IL-10 in supernatants from HIV-infected (In) and uninfected (Un) MDMs obtained from one representative healthy donor. (B) Data combined from six independent experiments (donors), each evaluated in triplicate. Here, data are shown as the ratio between cytokine concentrations found under the infection condition versus the uninfected counterpart. Cells were stimulated with MIF as follows: 0, 1, 10, or 25 ng/ml. Data shown in the gray boxes depict CD74 blocking (10 ng/ml of αCD74 or the corresponding isotype control) followed by MIF stimulation (1 or 25 ng/ml as denoted). Data represent the mean ± SD. Data were analyzed by one-way ANOVA followed by Tukey’s post-test. * p

    Article Snippet: Cytokine Quantitation The levels of the following cytokines were evaluated in MDM supernatants using commercially available ELISA sets: IL-8, IL-6, IL-1β, TNFα, IL-10 (ELISA MAX Deluxe kits, BioLegend) and sICAM (DouSet ELISA, R & D Systems).

    Techniques: Expressing, Migration, Infection, Derivative Assay, Blocking Assay

    Levels of proinflammatory cytokines released from S . Typhi- or S . Typhimurium-infected MDMs. Shown are levels of TNF-α (A) and IL-6 (B) released from MDMs after 2.5 and 24 h of S . Typhi infection and TNF-α (C) and IL-6 (D) released from MDMs after 2.5 and 24 h of S . Typhimurium infection. Statistical significance was determined by Student's t test comparing cytokine levels between LPS modification mutant-infected samples and the WT-infected sample collected at that time point. *, P

    Journal: Infection and Immunity

    Article Title: Cathelicidin Antimicrobial Peptide Expression Is Not Induced or Required for Bacterial Clearance during Salmonella enterica Infection of Human Monocyte-Derived Macrophages

    doi: 10.1128/IAI.00672-12

    Figure Lengend Snippet: Levels of proinflammatory cytokines released from S . Typhi- or S . Typhimurium-infected MDMs. Shown are levels of TNF-α (A) and IL-6 (B) released from MDMs after 2.5 and 24 h of S . Typhi infection and TNF-α (C) and IL-6 (D) released from MDMs after 2.5 and 24 h of S . Typhimurium infection. Statistical significance was determined by Student's t test comparing cytokine levels between LPS modification mutant-infected samples and the WT-infected sample collected at that time point. *, P

    Article Snippet: The proinflammatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) were detected in cell culture supernatant by enzyme-linked immunosorbent assay (ELISA) (Biolegend, San Diego, CA, and R & D Systems, Minneapolis, MN, respectively).

    Techniques: Infection, Modification, Mutagenesis

    DNA-PKcs is involved in the IL-6 and IL-12 response to CpG-ODN. Bone marrow-derived dendritic cells (DCs) from WT, DNA-PKcs −/− , TLR9 −/− and DNA-PKcs −/−/ TLR9 −/− mice were harvested based on their natural status: adhesion and suspension. ( A ). Combined DCs (adhesion and suspension) at day 7.5 were subjected to flow cytometry. The levels of CD11b, CD11c and MHC-II expression on DCs were determined. ( B–M ). Adhesion or suspension DCs at day 6.5 were seeded in 96-well plates at 1×10 5 /well in triplicate, and then treated with CpG-ODN (CpG, 0.2, 0.73, 2.2 or 7.3 µg/ml), LPS (0.1 µg/ml), R848 (0.125 µg/ml), or left untreated for indicated time durations. The levels of IL-6 ( B–G ) and IL-12p40 ( H–M ) were determined by ELISA using IL-6 or IL-12p40 ELISA kits. Similar results were obtained in at least 3 independent experiments. Note: bars represent the average of triplicates ± SD. * p

    Journal: PLoS ONE

    Article Title: Involvement of DNA-PKcs in the IL-6 and IL-12 Response to CpG-ODN Is Mediated by Its Interaction with TRAF6 in Dendritic Cells

    doi: 10.1371/journal.pone.0058072

    Figure Lengend Snippet: DNA-PKcs is involved in the IL-6 and IL-12 response to CpG-ODN. Bone marrow-derived dendritic cells (DCs) from WT, DNA-PKcs −/− , TLR9 −/− and DNA-PKcs −/−/ TLR9 −/− mice were harvested based on their natural status: adhesion and suspension. ( A ). Combined DCs (adhesion and suspension) at day 7.5 were subjected to flow cytometry. The levels of CD11b, CD11c and MHC-II expression on DCs were determined. ( B–M ). Adhesion or suspension DCs at day 6.5 were seeded in 96-well plates at 1×10 5 /well in triplicate, and then treated with CpG-ODN (CpG, 0.2, 0.73, 2.2 or 7.3 µg/ml), LPS (0.1 µg/ml), R848 (0.125 µg/ml), or left untreated for indicated time durations. The levels of IL-6 ( B–G ) and IL-12p40 ( H–M ) were determined by ELISA using IL-6 or IL-12p40 ELISA kits. Similar results were obtained in at least 3 independent experiments. Note: bars represent the average of triplicates ± SD. * p

    Article Snippet: The supernatants were collected and the production of IL-6 or IL-12p40 was determined by enzyme-linked immunosorbent assay (ELISA) kits (Biolegend) based on the manufacturer’s instruction.

    Techniques: Derivative Assay, Mouse Assay, Flow Cytometry, Cytometry, Expressing, Enzyme-linked Immunosorbent Assay