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  • 99
    Thermo Fisher tumor necrosis factor α tnf α
    Attenuation of the level of reactive oxygen species (ROS) by genistein. (A) Keratinocytes were stimulated respectively with a proinflammatory “cytokine mix” corresponding to a concentration of 2 ng/mL (ACT 2) or 5 ng/mL (ACT 5) in each compound of the mix and incubated with 10 mM N-acetyl-cysteine (NAC) (ACT 2 + NAC), 100 μM genistein (ACT 2 + GEN), 10 ng/mL <t>TNF-α</t> or 1 μg/mL LPS alone, and with 10 ng/mL TNF-α or 1 μg/mL LPS incubated with 100 μM genistein (TNF-α + GEN or LPS + GEN). DMSO-treated, unstimulated cells were used as the control (NACT). Additional control was used in the form of unstimulated cells treated with 100 μM genistein (GEN). Intracellular ROS levels were examined by a CellROX Deep Red Reagent and confocal fluorescence microscopy. Nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments (with scale bars 25 μm) are shown. (B) Analysis of ROS were additionally performed by fluorescent cell analyzer. The data are presented as the means ± standard deviation (SD) from three independent experiments. Significant differences ( p ≤ 0.05) between cell populations not expressing intracellular ROS (ROS [–]) and expressing intracellular ROS (ROS [+]) were observed for all tested conditions, except for LPS where cell numbers of ROS (-) and ROS (+) were comparable. Most important, statistically significant differences of p ≤ 0.05 within ROS groups are indicated with * for samples of TNF-α, TNF-α + GEN, and LPS versus NACT, with † for sample TNF-α + GEN with respect to TNF-α, while with ‡ for sample LPS + GEN referred to LPS. Statistical analysis was performed using ANOVA with Tukey’s HSD test.
    Tumor Necrosis Factor α Tnf α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 637 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
    Tumor Necrosis Factor α Tnf α, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 372 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
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    BioLegend tumour necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
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    Becton Dickinson tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
    Tumor Necrosis Factor α Tnf α, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 415 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
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    Cell Signaling Technology Inc tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
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    Haematologic Technologies tumor necrosis factor α tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
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    Abcam tumor necrosis factor α tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
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    Bio-Rad tumor necrosis factor α tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
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    USCN Life tumor necrosis factor α tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
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    Roche tumor necrosis factor α tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
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    ApexBio tumour necrosis factor α tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
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    Boehringer Mannheim tumor necrosis factor α tnf α
    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of <t>TNF-α,</t> IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P
    Tumor Necrosis Factor α Tnf α, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 89/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc tumor necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
    Tumor Necrosis Factor α Tnf α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cusabio tumor necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Elabscience tumor necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Proteintech tumor necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Boster Bio tumor necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    VECTOR-BEST tumour necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
    Tumour Necrosis Factor α Tnf α, supplied by VECTOR-BEST, used in various techniques. Bioz Stars score: 82/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GENTAUR tumor necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Meso Scale Diagnostics LLC tumor necrosis factor α tnf α
    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor <t>(TNF)-α</t> ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P
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    Image Search Results


    Attenuation of the level of reactive oxygen species (ROS) by genistein. (A) Keratinocytes were stimulated respectively with a proinflammatory “cytokine mix” corresponding to a concentration of 2 ng/mL (ACT 2) or 5 ng/mL (ACT 5) in each compound of the mix and incubated with 10 mM N-acetyl-cysteine (NAC) (ACT 2 + NAC), 100 μM genistein (ACT 2 + GEN), 10 ng/mL TNF-α or 1 μg/mL LPS alone, and with 10 ng/mL TNF-α or 1 μg/mL LPS incubated with 100 μM genistein (TNF-α + GEN or LPS + GEN). DMSO-treated, unstimulated cells were used as the control (NACT). Additional control was used in the form of unstimulated cells treated with 100 μM genistein (GEN). Intracellular ROS levels were examined by a CellROX Deep Red Reagent and confocal fluorescence microscopy. Nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments (with scale bars 25 μm) are shown. (B) Analysis of ROS were additionally performed by fluorescent cell analyzer. The data are presented as the means ± standard deviation (SD) from three independent experiments. Significant differences ( p ≤ 0.05) between cell populations not expressing intracellular ROS (ROS [–]) and expressing intracellular ROS (ROS [+]) were observed for all tested conditions, except for LPS where cell numbers of ROS (-) and ROS (+) were comparable. Most important, statistically significant differences of p ≤ 0.05 within ROS groups are indicated with * for samples of TNF-α, TNF-α + GEN, and LPS versus NACT, with † for sample TNF-α + GEN with respect to TNF-α, while with ‡ for sample LPS + GEN referred to LPS. Statistical analysis was performed using ANOVA with Tukey’s HSD test.

    Journal: PLoS ONE

    Article Title: Molecular action of isoflavone genistein in the human epithelial cell line HaCaT

    doi: 10.1371/journal.pone.0192297

    Figure Lengend Snippet: Attenuation of the level of reactive oxygen species (ROS) by genistein. (A) Keratinocytes were stimulated respectively with a proinflammatory “cytokine mix” corresponding to a concentration of 2 ng/mL (ACT 2) or 5 ng/mL (ACT 5) in each compound of the mix and incubated with 10 mM N-acetyl-cysteine (NAC) (ACT 2 + NAC), 100 μM genistein (ACT 2 + GEN), 10 ng/mL TNF-α or 1 μg/mL LPS alone, and with 10 ng/mL TNF-α or 1 μg/mL LPS incubated with 100 μM genistein (TNF-α + GEN or LPS + GEN). DMSO-treated, unstimulated cells were used as the control (NACT). Additional control was used in the form of unstimulated cells treated with 100 μM genistein (GEN). Intracellular ROS levels were examined by a CellROX Deep Red Reagent and confocal fluorescence microscopy. Nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments (with scale bars 25 μm) are shown. (B) Analysis of ROS were additionally performed by fluorescent cell analyzer. The data are presented as the means ± standard deviation (SD) from three independent experiments. Significant differences ( p ≤ 0.05) between cell populations not expressing intracellular ROS (ROS [–]) and expressing intracellular ROS (ROS [+]) were observed for all tested conditions, except for LPS where cell numbers of ROS (-) and ROS (+) were comparable. Most important, statistically significant differences of p ≤ 0.05 within ROS groups are indicated with * for samples of TNF-α, TNF-α + GEN, and LPS versus NACT, with † for sample TNF-α + GEN with respect to TNF-α, while with ‡ for sample LPS + GEN referred to LPS. Statistical analysis was performed using ANOVA with Tukey’s HSD test.

    Article Snippet: For further experiments, cells were stimulated with a combination of a proinflammatory “cytokine mix”: IL-1A, IL-17A, IL-22, oncostatin M (OSM), and tumor necrosis factor-α (TNF-α) (Gibco, Thermo Fisher Scientific, CA, USA) 2 ng/mL each or lipopolysaccharide (LPS, from Escherichia coli 055 :B5 , Sigma-Aldrich, St. Louis, USA) 1 μg/mL in the presence or absence of genistein for 24 hours [ ].

    Techniques: Concentration Assay, Activated Clotting Time Assay, Incubation, Fluorescence, Microscopy, Staining, Standard Deviation, Expressing

    TM5441 pre-treatment and siPAI-1 improved TNF-α-induced mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p

    Journal: Oncotarget

    Article Title: TM5441, a plasminogen activator inhibitor-1 inhibitor, protects against high fat diet-induced non-alcoholic fatty liver disease

    doi: 10.18632/oncotarget.21120

    Figure Lengend Snippet: TM5441 pre-treatment and siPAI-1 improved TNF-α-induced mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p

    Article Snippet: Growth arrested and synchronized cells were exposed with 50 ng/mL tumor necrosis factor-α/TNF-α (Sigma-Aldrich, St. Louis, MO, USA) or 25 and 50 nM mouse recombinant PAI-1 (EMD Millipore, Billerica, MA, USA) in the presence and absence of 20 μM TM5441.

    Techniques: Pyrolysis Gas Chromatography, Quantitative RT-PCR, Transfection, Expressing

    Early TM5441 treatment suppressed hepatic inflammation in HFD mice ( A , B ) Anti-inflammatory effect of TM5441 was analyzed through quantifying positive F4/80 IHC staining area in the liver section. Magnification, 200×; scale bar, 50 μm. Pro-inflammatory mRNA expressions in the liver, such as ( C ) F4/80, ( D ) MCP-1 ( E ) TNF-α, ( F ) NLRP3, were measured by RT-qPCR. Data are shown as mean ± SE of 7–8 mice. * p

    Journal: Oncotarget

    Article Title: TM5441, a plasminogen activator inhibitor-1 inhibitor, protects against high fat diet-induced non-alcoholic fatty liver disease

    doi: 10.18632/oncotarget.21120

    Figure Lengend Snippet: Early TM5441 treatment suppressed hepatic inflammation in HFD mice ( A , B ) Anti-inflammatory effect of TM5441 was analyzed through quantifying positive F4/80 IHC staining area in the liver section. Magnification, 200×; scale bar, 50 μm. Pro-inflammatory mRNA expressions in the liver, such as ( C ) F4/80, ( D ) MCP-1 ( E ) TNF-α, ( F ) NLRP3, were measured by RT-qPCR. Data are shown as mean ± SE of 7–8 mice. * p

    Article Snippet: Growth arrested and synchronized cells were exposed with 50 ng/mL tumor necrosis factor-α/TNF-α (Sigma-Aldrich, St. Louis, MO, USA) or 25 and 50 nM mouse recombinant PAI-1 (EMD Millipore, Billerica, MA, USA) in the presence and absence of 20 μM TM5441.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Quantitative RT-PCR

    β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of TNF-α, IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effect of β-patchoulene on cerebral ischemia-reperfusion injury and the TLR4/NF-κB signaling pathway

    doi: 10.3892/etm.2019.7374

    Figure Lengend Snippet: β-PAE reduces the inflammatory response and oxidative stress of rats with the I/R injury. (A) Western blotting was used to analyze the expression levels of TNF-α, IL-1β and IL-6 in the brain tissue. (B) The mRNA levels of TNF-α, IL-1β and IL-6 were measured using reverse transcription-quantitative polymerase chain reaction. (C) Superoxide flashes were measured to evaluate mitochondrial superoxide generation in brain tissues. (D) Oxidative damage parameters of brain cells, including MDA, GSH-Px and SOD levels were measured using the corresponding detection kits. Experiments were repeated at least three times, and data are presented as the mean ± standard deviation. *P

    Article Snippet: After blocking with 5% BSA (Bio-Rad Laboratories, Inc., Hercules, CA, USA) for 1 h at room temperature, membranes loaded with total proteins were incubated with the following antibodies: Rabbit anti-rat apoptosis regulator BAX (Bax; cat. no. 14796; 1:1,000), apoptosis regulator Bcl-2 (Bcl-2; cat. no. 3498; 1:1,000), TNF-α (cat. no. 8184; 1:1,000), IL-1β (cat. no. 12703; 1:1,000), IL-6 (cat. no. 12912; 1:1,000), TLR4 (cat. no. 14358; 1:1,000), IκBα (cat. no. 4812; 1:1,000), β-actin (cat. no. 4970; 1:1,000; all Cell Signaling Technology, Inc., Danvers, MA, USA) overnight at 4°C.

    Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Multiple Displacement Amplification, Standard Deviation

    Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor (TNF)-α ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Inhibition of Epidermal Growth Factor Receptor (EGFR) Reduces Lipopolysaccharide (LPS)-Induced Activation and Inflammatory Cytokines in Hepatic Stellate Cells In Vitro

    doi: 10.12659/MSM.909901

    Figure Lengend Snippet: Knockdown of the EGFR gene following small interfering RNA (siRNA) transfection reduced lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) injury. ( A ) Western blot analysis of epidermal growth factor receptor (EGFR) following small interfering RNA (siRNA) transfection in HSCs (NC – negative control for transfection). After incubation of the transfected hepatic stellate cells (HSCs) for 24 h, EGFR gene knockdown HSCs were stimulated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times (Si – EGFR siRNA). ( B, C ) HSCs incubated with LPS for 24 h. Tumor necrosis factor (TNF)-α ( B ) and interleukin (IL)-6 ( C ) in cell lysates were detected using Western blot. ( D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α and IL-6 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized by β-actin. ( E ) HSCs incubated with LPS for 1 h. IκB-α, cytoplasm NFκB (C-NFκB), and nuclear NFκB (N-NFκB) protein levels were detected using Western blot. ( F ) HSCs incubated with LPS for 24 h. The levels of transforming growth factor (TGF)-β, Col-1, and α-smooth muscle actin (SMA) were detected using Western blot. ( G ) HSCs incubated with LPS for 6 h. The mRNA levels of TGF-β, Col-1, and α-SMA were detected by RT-qPCR and normalized against β-actin. * P

    Article Snippet: Primary antibodies to epidermal growth factor receptor (EGFR), phospho-EGFR (p-EGFR), Tyr835, TLR4, tumor necrosis factor (TNF)-α, interleukin (IL)-6, IκB-α (an inhibitor of NF-κB) and NFκB P65 were obtained from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Small Interfering RNA, Transfection, Western Blot, Negative Control, Incubation, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    Treatment with AG1478 reduced levels of lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) inflammatory cytokines. Hepatic stellate cells (HSCs) were pretreated with AG1478 (2.5 μM and 5 μM) for 2 h, and then exposed to lipopolysaccharide (LPS) (100 ng/mL) for the indicated times. ( A, B ) HSCs incubated with LPS for 24 h. Levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6, in the cell lysates were detected by Western blot ( A ). The figures in the columns show the normalized optical density (OD) for the data from three independent experiments ( B ). ( C, D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α ( C ) and IL-6 ( D ) were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized against β-actin. ( E–G ) Western blot analysis of IκBα ( E ), cytoplasm NFκB P65 (C-NFκB) ( F ) and nuclear NFκB P65(N-NFκB) ( G ) levels in HSCs incubated with LPS for 1h. GAPDH was used as a loading control for IκBα/C-NFκB and laminin B as loading control for N-NFκB. ns – not significant vs. Ctrl group; ** P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Inhibition of Epidermal Growth Factor Receptor (EGFR) Reduces Lipopolysaccharide (LPS)-Induced Activation and Inflammatory Cytokines in Hepatic Stellate Cells In Vitro

    doi: 10.12659/MSM.909901

    Figure Lengend Snippet: Treatment with AG1478 reduced levels of lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) inflammatory cytokines. Hepatic stellate cells (HSCs) were pretreated with AG1478 (2.5 μM and 5 μM) for 2 h, and then exposed to lipopolysaccharide (LPS) (100 ng/mL) for the indicated times. ( A, B ) HSCs incubated with LPS for 24 h. Levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6, in the cell lysates were detected by Western blot ( A ). The figures in the columns show the normalized optical density (OD) for the data from three independent experiments ( B ). ( C, D ) HSCs incubated with LPS for 6 h. The mRNA levels of TNF-α ( C ) and IL-6 ( D ) were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and normalized against β-actin. ( E–G ) Western blot analysis of IκBα ( E ), cytoplasm NFκB P65 (C-NFκB) ( F ) and nuclear NFκB P65(N-NFκB) ( G ) levels in HSCs incubated with LPS for 1h. GAPDH was used as a loading control for IκBα/C-NFκB and laminin B as loading control for N-NFκB. ns – not significant vs. Ctrl group; ** P

    Article Snippet: Primary antibodies to epidermal growth factor receptor (EGFR), phospho-EGFR (p-EGFR), Tyr835, TLR4, tumor necrosis factor (TNF)-α, interleukin (IL)-6, IκB-α (an inhibitor of NF-κB) and NFκB P65 were obtained from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    Effects of doxycycline on expression of TNF-α protein by Western blotting analysis and ELISA. (A) HaCaT-TNF-α (TNF-α) and HaCaT-GFP (GFP) cells were treated with increasing doxycycline (Dox) concentrations (0, 0.5, 1.0, 2.0, and 3.0 μ g/mL) for 48 hrs. Cells were lyzed and analyzed using Western blot with TNF-α-specific antibody rabbit monoclonal antibody. Fifteen micrograms of proteins were loaded on each lane. GAPDH was used as loading control. Bar graphs represent relative TNF-α expression normalized with GAPDH. (B) Same as (A) except HaCaT-TNF-α and HaCaT-GFP cells were treated with 1.0 μ g/ml Dox and incubated for 24, 48 and 72 hrs. (C) HaCaT-TNF-α were treated with increasing doses of Dox (1.0, 2.0, and 3.0 μ g/mL) and incubated for 24, 48 and 72 hrs to induce secretion of TNF-α proteins. The supernatants were prepared and detected by ELISA. Line graphs represent concentration of TNF-α secreted into the supernatant after treated with doxycycline. All data represent mean ± SEM from three independent experiments. (D) HaCaT-TNF-α were treated with and without 1.0 μ g/ml Dox and incubated for 48. Cells were harvested and subjected to subcellular fractionation into cytoplasmic (Cyt) and membrane (Mem) fractions. Fifteen micrograms of proteins were loaded on each lane. GAPDH was used as cytoplasmic markers and loading control. Bar graphs represent relative TNF-α expression normalized with GAPDH. ** (p

    Journal: PLoS ONE

    Article Title: Assessment of Anti-TNF-α Activities in Keratinocytes Expressing Inducible TNF- α: A Novel Tool for Anti-TNF-α Drug Screening

    doi: 10.1371/journal.pone.0159151

    Figure Lengend Snippet: Effects of doxycycline on expression of TNF-α protein by Western blotting analysis and ELISA. (A) HaCaT-TNF-α (TNF-α) and HaCaT-GFP (GFP) cells were treated with increasing doxycycline (Dox) concentrations (0, 0.5, 1.0, 2.0, and 3.0 μ g/mL) for 48 hrs. Cells were lyzed and analyzed using Western blot with TNF-α-specific antibody rabbit monoclonal antibody. Fifteen micrograms of proteins were loaded on each lane. GAPDH was used as loading control. Bar graphs represent relative TNF-α expression normalized with GAPDH. (B) Same as (A) except HaCaT-TNF-α and HaCaT-GFP cells were treated with 1.0 μ g/ml Dox and incubated for 24, 48 and 72 hrs. (C) HaCaT-TNF-α were treated with increasing doses of Dox (1.0, 2.0, and 3.0 μ g/mL) and incubated for 24, 48 and 72 hrs to induce secretion of TNF-α proteins. The supernatants were prepared and detected by ELISA. Line graphs represent concentration of TNF-α secreted into the supernatant after treated with doxycycline. All data represent mean ± SEM from three independent experiments. (D) HaCaT-TNF-α were treated with and without 1.0 μ g/ml Dox and incubated for 48. Cells were harvested and subjected to subcellular fractionation into cytoplasmic (Cyt) and membrane (Mem) fractions. Fifteen micrograms of proteins were loaded on each lane. GAPDH was used as cytoplasmic markers and loading control. Bar graphs represent relative TNF-α expression normalized with GAPDH. ** (p

    Article Snippet: Both membrane-bound and secreted TNF-α are biologically active and there is evidence to suggest membrane-bound TNF-α also plays a role in cell-cell contact mediated juxtacrine cell signaling [ ].

    Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Concentration Assay, Fractionation

    Quercetin inhibits pro-inflammatory cytokine (IL-1β and IL-8) expression. HaCaT-TNF-α cells pre-treated with 1.0 μ g/mL Dox or 10 ng/mL TNF-α for 24 hrs. Cells were then treated with 10 μ M and 20 μ M Quercetin for an additional 24 hrs. Total RNA was extracted and reverse-transcribed. cDNA was amplified by qRT-PCR. Relative expression of IL-1β (A-B) and IL-8 expression (C-D) are shown. Results are represented as Mean ± SEM from three independent experiments compared with control # ( p

    Journal: PLoS ONE

    Article Title: Assessment of Anti-TNF-α Activities in Keratinocytes Expressing Inducible TNF- α: A Novel Tool for Anti-TNF-α Drug Screening

    doi: 10.1371/journal.pone.0159151

    Figure Lengend Snippet: Quercetin inhibits pro-inflammatory cytokine (IL-1β and IL-8) expression. HaCaT-TNF-α cells pre-treated with 1.0 μ g/mL Dox or 10 ng/mL TNF-α for 24 hrs. Cells were then treated with 10 μ M and 20 μ M Quercetin for an additional 24 hrs. Total RNA was extracted and reverse-transcribed. cDNA was amplified by qRT-PCR. Relative expression of IL-1β (A-B) and IL-8 expression (C-D) are shown. Results are represented as Mean ± SEM from three independent experiments compared with control # ( p

    Article Snippet: Both membrane-bound and secreted TNF-α are biologically active and there is evidence to suggest membrane-bound TNF-α also plays a role in cell-cell contact mediated juxtacrine cell signaling [ ].

    Techniques: Expressing, Amplification, Quantitative RT-PCR

    Effects of doxycycline induces pro-inflamatory cytokines (IL-1β, IL-8, IL-6, NF-κB1, KRT16, FOSL1 and MMP9) expression in HaCaT-TNF-α cells. HaCaT-TNF-α cells treated with 1.0 μ g/mL Dox or 10 ng/mL TNF-α for 24 hrs were then treated with or without 10 ng/mL IFN-γ for a further 24 hrs. Total RNA was extracted and reverse-transcribed. cDNA was amplified by qRT-PCR using IL-1β, IL-8, IL-6, NF-κB1, KRT16, FOSL1 and MMP9 primers ( Table 1 ). GAPDH was used as loading control. Relative expression of IL-1β (A) IL-8 (B) IL-6 (C) NF-κB1 (D) KRT16 (E) FOSL1 (F) and MMP9 expression (G) are shown. Values represent relative gene expression normalized with GAPDH. Mean ± SEM from three independent experiments. *( p

    Journal: PLoS ONE

    Article Title: Assessment of Anti-TNF-α Activities in Keratinocytes Expressing Inducible TNF- α: A Novel Tool for Anti-TNF-α Drug Screening

    doi: 10.1371/journal.pone.0159151

    Figure Lengend Snippet: Effects of doxycycline induces pro-inflamatory cytokines (IL-1β, IL-8, IL-6, NF-κB1, KRT16, FOSL1 and MMP9) expression in HaCaT-TNF-α cells. HaCaT-TNF-α cells treated with 1.0 μ g/mL Dox or 10 ng/mL TNF-α for 24 hrs were then treated with or without 10 ng/mL IFN-γ for a further 24 hrs. Total RNA was extracted and reverse-transcribed. cDNA was amplified by qRT-PCR using IL-1β, IL-8, IL-6, NF-κB1, KRT16, FOSL1 and MMP9 primers ( Table 1 ). GAPDH was used as loading control. Relative expression of IL-1β (A) IL-8 (B) IL-6 (C) NF-κB1 (D) KRT16 (E) FOSL1 (F) and MMP9 expression (G) are shown. Values represent relative gene expression normalized with GAPDH. Mean ± SEM from three independent experiments. *( p

    Article Snippet: Both membrane-bound and secreted TNF-α are biologically active and there is evidence to suggest membrane-bound TNF-α also plays a role in cell-cell contact mediated juxtacrine cell signaling [ ].

    Techniques: Expressing, Amplification, Quantitative RT-PCR

    Effects of TNF-α expression on HaCaT cell viabilities. Line graphs represent percent cell viability of HaCaT-GFP (A) HaCaT-TNF-α (B) after treated with doxycycline (1.0, 2.0 and 3.0 μ g/mL) and 10 ng/mL TNF-α was used as a positive control. Values are represented as mean ± SEM (n = 3). (Percentage cell viability is shown as a percent of doxycycline-treated cells normalized with control untreated cells.)

    Journal: PLoS ONE

    Article Title: Assessment of Anti-TNF-α Activities in Keratinocytes Expressing Inducible TNF- α: A Novel Tool for Anti-TNF-α Drug Screening

    doi: 10.1371/journal.pone.0159151

    Figure Lengend Snippet: Effects of TNF-α expression on HaCaT cell viabilities. Line graphs represent percent cell viability of HaCaT-GFP (A) HaCaT-TNF-α (B) after treated with doxycycline (1.0, 2.0 and 3.0 μ g/mL) and 10 ng/mL TNF-α was used as a positive control. Values are represented as mean ± SEM (n = 3). (Percentage cell viability is shown as a percent of doxycycline-treated cells normalized with control untreated cells.)

    Article Snippet: Both membrane-bound and secreted TNF-α are biologically active and there is evidence to suggest membrane-bound TNF-α also plays a role in cell-cell contact mediated juxtacrine cell signaling [ ].

    Techniques: Expressing, Positive Control

    Effects of doxycycline induces pro-inflamatory cytokines (IL-1β and IL-8) expression in HaCaT cells. HaCaT cells were treated with 1.0 μ g/mL doxycycline (Dox) or 10 ng/mL TNF-α for 24 hrs. and then treated with or without 10 ng/mL IFN-γ for a further 24 hrs. Total RNA was extracted and reverse-transcribed. cDNA was amplified by qRT-PCR using IL-1β and IL-8 primers ( Table 1 ). GAPDH was used as loading control. Bar graphs represent relative gene expression (A) IL-1β (B) IL-8 expression where values represent relative gene expression normalized with GAPDH. Mean ± SEM from three independent experiments. *( p

    Journal: PLoS ONE

    Article Title: Assessment of Anti-TNF-α Activities in Keratinocytes Expressing Inducible TNF- α: A Novel Tool for Anti-TNF-α Drug Screening

    doi: 10.1371/journal.pone.0159151

    Figure Lengend Snippet: Effects of doxycycline induces pro-inflamatory cytokines (IL-1β and IL-8) expression in HaCaT cells. HaCaT cells were treated with 1.0 μ g/mL doxycycline (Dox) or 10 ng/mL TNF-α for 24 hrs. and then treated with or without 10 ng/mL IFN-γ for a further 24 hrs. Total RNA was extracted and reverse-transcribed. cDNA was amplified by qRT-PCR using IL-1β and IL-8 primers ( Table 1 ). GAPDH was used as loading control. Bar graphs represent relative gene expression (A) IL-1β (B) IL-8 expression where values represent relative gene expression normalized with GAPDH. Mean ± SEM from three independent experiments. *( p

    Article Snippet: Both membrane-bound and secreted TNF-α are biologically active and there is evidence to suggest membrane-bound TNF-α also plays a role in cell-cell contact mediated juxtacrine cell signaling [ ].

    Techniques: Expressing, Amplification, Quantitative RT-PCR

    Effects of secreted TNF-α in supernatant stimulated pro-inflammatory cytokines (IL-1β and IL-8) expression and effects of membrane TNF-α on IL-8 expression in HaCaT cells. HaCaT-TNF-α cells were treated with 1.0 μ g/mL doxycycline (Dox) for 48 hrs. Supernatant collected was serially diluted with fresh medium in the ratio (supernatant fresh medium) 1:1.5, 1:2, 1:3 and 1:4. HaCaT cells were incubated with diluted supernatant for 24 hrs and subsequently treated with 10 ng/mL IFN-γ for a further 24 hrs. Expression levels of IL-1β and IL-8 were quantitated by qRT-PCR. Relative expression of IL-1β (A) and IL-8 (B) are shown. (C) HaCaT-TNF-α cells were cultured at different seeding densities and treated with 1 μ g/ml Dox for 48 hr. Cells harvested at approximately 50% or 90% confluence. IL-8 gene expression from cells at 50% and 90% confluency were analyzed by RT qPCR. Relative expression of IL-8 are shown. Results are represented as the Mean ± SEM from three independent experiments compared with control untreated *( p

    Journal: PLoS ONE

    Article Title: Assessment of Anti-TNF-α Activities in Keratinocytes Expressing Inducible TNF- α: A Novel Tool for Anti-TNF-α Drug Screening

    doi: 10.1371/journal.pone.0159151

    Figure Lengend Snippet: Effects of secreted TNF-α in supernatant stimulated pro-inflammatory cytokines (IL-1β and IL-8) expression and effects of membrane TNF-α on IL-8 expression in HaCaT cells. HaCaT-TNF-α cells were treated with 1.0 μ g/mL doxycycline (Dox) for 48 hrs. Supernatant collected was serially diluted with fresh medium in the ratio (supernatant fresh medium) 1:1.5, 1:2, 1:3 and 1:4. HaCaT cells were incubated with diluted supernatant for 24 hrs and subsequently treated with 10 ng/mL IFN-γ for a further 24 hrs. Expression levels of IL-1β and IL-8 were quantitated by qRT-PCR. Relative expression of IL-1β (A) and IL-8 (B) are shown. (C) HaCaT-TNF-α cells were cultured at different seeding densities and treated with 1 μ g/ml Dox for 48 hr. Cells harvested at approximately 50% or 90% confluence. IL-8 gene expression from cells at 50% and 90% confluency were analyzed by RT qPCR. Relative expression of IL-8 are shown. Results are represented as the Mean ± SEM from three independent experiments compared with control untreated *( p

    Article Snippet: Both membrane-bound and secreted TNF-α are biologically active and there is evidence to suggest membrane-bound TNF-α also plays a role in cell-cell contact mediated juxtacrine cell signaling [ ].

    Techniques: Expressing, Incubation, Quantitative RT-PCR, Cell Culture

    The expression of TNF-α, CD3, CD4 and CD8 in periapical lesions. Inflammatory response of healthy oral mucosa (n=3), periapical granuloma (n=14) and radicular cyst (n=13) was detected by immunohistochemical staining. A, F, K. HE staining of healthy oral mucosa, periapical granuloma and radicular cyst. B, G, L. TNF-α significantly overexpressed in periapical granuloma, and mildly decreased in radicular cyst. In the healthy oral mucosa, TNF-α were scarcely detected. C, H, M. The infiltration of total T cells was detected by CD3 immunostaining. Similar to the expression of TNF-α, CD3 + T cells accumulated in granuloma and reduced in radicular cyst, while it is seldom observed in healthy oral mucosa. D, I, N, E, J, O. CD8 and CD4 were stained, the positive cells accumulated within periapical granuloma and radicular cyst, while CD8 + and CD4 + cells in healthy mucosa were barely detected. Scale bar = 50 μm.

    Journal: American Journal of Translational Research

    Article Title: The expression of PD-1 and LAG-3 in periapical lesions

    doi:

    Figure Lengend Snippet: The expression of TNF-α, CD3, CD4 and CD8 in periapical lesions. Inflammatory response of healthy oral mucosa (n=3), periapical granuloma (n=14) and radicular cyst (n=13) was detected by immunohistochemical staining. A, F, K. HE staining of healthy oral mucosa, periapical granuloma and radicular cyst. B, G, L. TNF-α significantly overexpressed in periapical granuloma, and mildly decreased in radicular cyst. In the healthy oral mucosa, TNF-α were scarcely detected. C, H, M. The infiltration of total T cells was detected by CD3 immunostaining. Similar to the expression of TNF-α, CD3 + T cells accumulated in granuloma and reduced in radicular cyst, while it is seldom observed in healthy oral mucosa. D, I, N, E, J, O. CD8 and CD4 were stained, the positive cells accumulated within periapical granuloma and radicular cyst, while CD8 + and CD4 + cells in healthy mucosa were barely detected. Scale bar = 50 μm.

    Article Snippet: Briefly, sections were incubated with first antibodies overnight at 4°C.The antibodies included: anti-CD4 (Abcam, Cambridge, MA, USA), anti-CD8 (Abcam, Cambridge, MA, USA), anti-CD3 (Abcam, Cambridge, MA, USA), anti-LAG-3 (Cell Signaling Technology, Danvers, MA, USA), anti-tumor necrosis factor (TNF)-α (Cell Signaling Technology), anti-PD-1 (Cell Signaling Technology).

    Techniques: Expressing, Immunohistochemistry, Staining, Immunostaining