il-1β Search Results


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  • 95
    Thermo Fisher recombinant interleukin 1β il 1β
    Upregulation of FSTL1 expression induced by inflammatory mediators in fibroblast-like synoviocytes (FLSs) from patients with RA . (A) RA FLSs ( n = 6) were treated with the indicated stimuli for 24 hours or were left untreated (medium). FSTL1 mRNA was determined using RT-PCR. The results are presented in relative units, and the P values derived from paired t -tests are indicated for each comparison. (B) FLSs from two RA patients (RA1 and RA2) were stimulated by combined <t>IL-1β</t> and TNFα or by TGFβ, or they were left untreated for 36 hours or 72 hours. Cell lysates were collected and analyzed by Western blot analysis using the anti-FSTL1 polyclonal antibody. β-Actin was used as a loading control. FSTL1, follistatin-like protein 1; RA, rheumatoid arthritis; PCR, polymerase chain reaction; IL-1β, <t>interleukin-1β;</t> TNFα, tumor necrosis factor α; TGFβ, transforming growth factor β; LPS, lipopolysaccharide; bLP, bacterial lipoprotein; PIC, polyinosinic:polycytidylic acid.
    Recombinant Interleukin 1β Il 1β, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore interleukin 1β il 1β
    Upregulation of FSTL1 expression induced by inflammatory mediators in fibroblast-like synoviocytes (FLSs) from patients with RA . (A) RA FLSs ( n = 6) were treated with the indicated stimuli for 24 hours or were left untreated (medium). FSTL1 mRNA was determined using RT-PCR. The results are presented in relative units, and the P values derived from paired t -tests are indicated for each comparison. (B) FLSs from two RA patients (RA1 and RA2) were stimulated by combined <t>IL-1β</t> and TNFα or by TGFβ, or they were left untreated for 36 hours or 72 hours. Cell lysates were collected and analyzed by Western blot analysis using the anti-FSTL1 polyclonal antibody. β-Actin was used as a loading control. FSTL1, follistatin-like protein 1; RA, rheumatoid arthritis; PCR, polymerase chain reaction; IL-1β, <t>interleukin-1β;</t> TNFα, tumor necrosis factor α; TGFβ, transforming growth factor β; LPS, lipopolysaccharide; bLP, bacterial lipoprotein; PIC, polyinosinic:polycytidylic acid.
    Interleukin 1β Il 1β, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boster Bio interleukin 1β il 1β
    Upregulation of FSTL1 expression induced by inflammatory mediators in fibroblast-like synoviocytes (FLSs) from patients with RA . (A) RA FLSs ( n = 6) were treated with the indicated stimuli for 24 hours or were left untreated (medium). FSTL1 mRNA was determined using RT-PCR. The results are presented in relative units, and the P values derived from paired t -tests are indicated for each comparison. (B) FLSs from two RA patients (RA1 and RA2) were stimulated by combined <t>IL-1β</t> and TNFα or by TGFβ, or they were left untreated for 36 hours or 72 hours. Cell lysates were collected and analyzed by Western blot analysis using the anti-FSTL1 polyclonal antibody. β-Actin was used as a loading control. FSTL1, follistatin-like protein 1; RA, rheumatoid arthritis; PCR, polymerase chain reaction; IL-1β, <t>interleukin-1β;</t> TNFα, tumor necrosis factor α; TGFβ, transforming growth factor β; LPS, lipopolysaccharide; bLP, bacterial lipoprotein; PIC, polyinosinic:polycytidylic acid.
    Interleukin 1β Il 1β, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems interleukin 1β il 1β
    Mindin mRNA expression is upregulated by CpG-ODN stimulation. About 1 × 10 5 RAW 264.7 cells were cultured in 12-well plates and stimulated with cytokines of mouse recombinant tumor necrosis factor-α (TNF-α) and <t>interleukin-1β</t> (IL-1β), toll-like receptor (TLR) ligands of Pam3-CSK4, peptidoglycans (PGN), lipopolysaccharide (LPS), flagellin and CpG-ODN 1585 (B) for 8 h, then cells were harvested for RNA isolation and relative mRNA expression of mindin was analyzed using quantitative real time polymerase chain reaction (RT-PCR). In each group, bars represent mean ± SD; b P
    Interleukin 1β Il 1β, supplied by R&D Systems, used in various techniques. Bioz Stars score: 89/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher interleukin 1β il 1β
    Mindin mRNA expression is upregulated by CpG-ODN stimulation. About 1 × 10 5 RAW 264.7 cells were cultured in 12-well plates and stimulated with cytokines of mouse recombinant tumor necrosis factor-α (TNF-α) and <t>interleukin-1β</t> (IL-1β), toll-like receptor (TLR) ligands of Pam3-CSK4, peptidoglycans (PGN), lipopolysaccharide (LPS), flagellin and CpG-ODN 1585 (B) for 8 h, then cells were harvested for RNA isolation and relative mRNA expression of mindin was analyzed using quantitative real time polymerase chain reaction (RT-PCR). In each group, bars represent mean ± SD; b P
    Interleukin 1β Il 1β, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    USCN Life interleukin 1β il 1β
    Mindin mRNA expression is upregulated by CpG-ODN stimulation. About 1 × 10 5 RAW 264.7 cells were cultured in 12-well plates and stimulated with cytokines of mouse recombinant tumor necrosis factor-α (TNF-α) and <t>interleukin-1β</t> (IL-1β), toll-like receptor (TLR) ligands of Pam3-CSK4, peptidoglycans (PGN), lipopolysaccharide (LPS), flagellin and CpG-ODN 1585 (B) for 8 h, then cells were harvested for RNA isolation and relative mRNA expression of mindin was analyzed using quantitative real time polymerase chain reaction (RT-PCR). In each group, bars represent mean ± SD; b P
    Interleukin 1β Il 1β, supplied by USCN Life, used in various techniques. Bioz Stars score: 89/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam interleukin 1β il 1β
    Mindin mRNA expression is upregulated by CpG-ODN stimulation. About 1 × 10 5 RAW 264.7 cells were cultured in 12-well plates and stimulated with cytokines of mouse recombinant tumor necrosis factor-α (TNF-α) and <t>interleukin-1β</t> (IL-1β), toll-like receptor (TLR) ligands of Pam3-CSK4, peptidoglycans (PGN), lipopolysaccharide (LPS), flagellin and CpG-ODN 1585 (B) for 8 h, then cells were harvested for RNA isolation and relative mRNA expression of mindin was analyzed using quantitative real time polymerase chain reaction (RT-PCR). In each group, bars represent mean ± SD; b P
    Interleukin 1β Il 1β, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quanterix il1β
    Concentrations of biomarkers for adults with DS with and without clinical dementia. a Aβ 40 , b Aβ 42 , c Aβ 42 /Aβ 40 , d Aβ 42 /t-tau, e t-tau, f <t>IL1β,</t> g IL10, h IL6, i TNFα, and j NfL. Lines indicate median value (pg/ml)
    Il1β, supplied by Quanterix, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioVision interleukin 1β il 1β
    Concentrations of biomarkers for adults with DS with and without clinical dementia. a Aβ 40 , b Aβ 42 , c Aβ 42 /Aβ 40 , d Aβ 42 /t-tau, e t-tau, f <t>IL1β,</t> g IL10, h IL6, i TNFα, and j NfL. Lines indicate median value (pg/ml)
    Interleukin 1β Il 1β, supplied by BioVision, used in various techniques. Bioz Stars score: 89/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti interleukin 1β il 1β
    Concentrations of biomarkers for adults with DS with and without clinical dementia. a Aβ 40 , b Aβ 42 , c Aβ 42 /Aβ 40 , d Aβ 42 /t-tau, e t-tau, f <t>IL1β,</t> g IL10, h IL6, i TNFα, and j NfL. Lines indicate median value (pg/ml)
    Anti Interleukin 1β Il 1β, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abnova interleukin 1β il 1β
    The mRNA abundance of <t>interleukin-1β</t> (IL-1β ), tumor necrosis factor-α (TNF-α ), IL-8 and xanthine oxidoreductase ( XOR ) in udder of rats. The mRNA abundances of IL-1β , TNF-α , IL-8 and XOR were determined by RT-PCR with mammary tissues obtained after saline infusion at day 14 of gestation from pregnant rats ( n = 7) and age-matched non-pregnant rats ( n = 11). Values are presented as means ± SE. Differences between pregnant and non-pregnant rats were indicated by asterisks (**** P
    Interleukin 1β Il 1β, supplied by Abnova, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore interleukin 1β il 1β levels
    The mRNA abundance of <t>interleukin-1β</t> (IL-1β ), tumor necrosis factor-α (TNF-α ), IL-8 and xanthine oxidoreductase ( XOR ) in udder of rats. The mRNA abundances of IL-1β , TNF-α , IL-8 and XOR were determined by RT-PCR with mammary tissues obtained after saline infusion at day 14 of gestation from pregnant rats ( n = 7) and age-matched non-pregnant rats ( n = 11). Values are presented as means ± SE. Differences between pregnant and non-pregnant rats were indicated by asterisks (**** P
    Interleukin 1β Il 1β Levels, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc il 1β
    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and <t>IL‐1β‐induced</t> increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p
    Il 1β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1329 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant interleukin 1β il 1β
    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and <t>IL‐1β‐induced</t> increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p
    Recombinant Interleukin 1β Il 1β, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human interleukin 1β il 1β
    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and <t>IL‐1β‐induced</t> increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p
    Recombinant Human Interleukin 1β Il 1β, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ImmunoTools interleukin 1β il 1β
    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and <t>IL‐1β‐induced</t> increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p
    Interleukin 1β Il 1β, supplied by ImmunoTools, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems interleukin 1β il 1β elisa kit
    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and <t>IL‐1β‐induced</t> increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p
    Interleukin 1β Il 1β Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA interleukin 1β il 1β
    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and <t>IL‐1β‐induced</t> increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p
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    Meso Scale Diagnostics LLC interleukin 1β il 1β
    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and <t>IL‐1β‐induced</t> increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p
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    Santa Cruz Biotechnology interleukin 1β il 1β
    Cytosolic adaptor caspase recruitment domain 9 (CARD9) deficiency inhibits angiotensin II (Ang II) infusion–induced proinflammatory cell infiltration and expression of inflammatory cytokines. ( a ) Heart sections from wild-type (WT) and CARD9 −/− mice treated with saline or Ang II at 1,500 ng/kg/min for 7 days were stained with hematoxylin and eosin (HE). ( b – d ) Heart sections from WT and CARD9 −/− mice were detected by immunohistochemistry with antibodies against Mac-2, <t>interleukin-1β</t> (IL-1β), and connective tissue growth factor (CTGF) (top). Bar graph shows quantified areas (bottom). Bar = 50 µm. Data represent mean ± s.e.m. ( n = 5). * P
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    PeproTech interleukin 1β il 1β
    Cytosolic adaptor caspase recruitment domain 9 (CARD9) deficiency inhibits angiotensin II (Ang II) infusion–induced proinflammatory cell infiltration and expression of inflammatory cytokines. ( a ) Heart sections from wild-type (WT) and CARD9 −/− mice treated with saline or Ang II at 1,500 ng/kg/min for 7 days were stained with hematoxylin and eosin (HE). ( b – d ) Heart sections from WT and CARD9 −/− mice were detected by immunohistochemistry with antibodies against Mac-2, <t>interleukin-1β</t> (IL-1β), and connective tissue growth factor (CTGF) (top). Bar graph shows quantified areas (bottom). Bar = 50 µm. Data represent mean ± s.e.m. ( n = 5). * P
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    Millipore recombinant interleukin 1β il 1β
    Catabolic regulation of LRP5 mediates via β-catenin-Tcf/Lef signaling. (A) Chondrocytes were transfected with 1 μg of empty vector (EV) or pSPORT6- Lrp5 plus the TOPflash or FOPflash reporter constructs. After 24 hours, transcriptional activation by β-catenin was determined by luciferase reporter gene assays ( n = 7 independent experiments). (B) Chondrocytes obtained from wild-type (WT) and Lrp5 -/- mice were treated with 1 ng/ml <t>interleukin</t> <t>1β</t> (IL-1β), 50 ng/ml Wnt3a or 500 ng/ml Wnt7a, and transcriptional activation by β-catenin was evaluated by luciferase reporter gene assays ( n = 6). Values are expressed as means ± SEM (* P
    Recombinant Interleukin 1β Il 1β, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc il 1β antibody
    PmpG-1-vaults activate the NLRP3 inflammasome and induce <t>IL-1β</t> secretion, as measured by an ELISA assay
    Il 1β Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Upregulation of FSTL1 expression induced by inflammatory mediators in fibroblast-like synoviocytes (FLSs) from patients with RA . (A) RA FLSs ( n = 6) were treated with the indicated stimuli for 24 hours or were left untreated (medium). FSTL1 mRNA was determined using RT-PCR. The results are presented in relative units, and the P values derived from paired t -tests are indicated for each comparison. (B) FLSs from two RA patients (RA1 and RA2) were stimulated by combined IL-1β and TNFα or by TGFβ, or they were left untreated for 36 hours or 72 hours. Cell lysates were collected and analyzed by Western blot analysis using the anti-FSTL1 polyclonal antibody. β-Actin was used as a loading control. FSTL1, follistatin-like protein 1; RA, rheumatoid arthritis; PCR, polymerase chain reaction; IL-1β, interleukin-1β; TNFα, tumor necrosis factor α; TGFβ, transforming growth factor β; LPS, lipopolysaccharide; bLP, bacterial lipoprotein; PIC, polyinosinic:polycytidylic acid.

    Journal: Arthritis Research & Therapy

    Article Title: Follistatin-like protein 1 is elevated in systemic autoimmune diseases and correlated with disease activity in patients with rheumatoid arthritis

    doi: 10.1186/ar3241

    Figure Lengend Snippet: Upregulation of FSTL1 expression induced by inflammatory mediators in fibroblast-like synoviocytes (FLSs) from patients with RA . (A) RA FLSs ( n = 6) were treated with the indicated stimuli for 24 hours or were left untreated (medium). FSTL1 mRNA was determined using RT-PCR. The results are presented in relative units, and the P values derived from paired t -tests are indicated for each comparison. (B) FLSs from two RA patients (RA1 and RA2) were stimulated by combined IL-1β and TNFα or by TGFβ, or they were left untreated for 36 hours or 72 hours. Cell lysates were collected and analyzed by Western blot analysis using the anti-FSTL1 polyclonal antibody. β-Actin was used as a loading control. FSTL1, follistatin-like protein 1; RA, rheumatoid arthritis; PCR, polymerase chain reaction; IL-1β, interleukin-1β; TNFα, tumor necrosis factor α; TGFβ, transforming growth factor β; LPS, lipopolysaccharide; bLP, bacterial lipoprotein; PIC, polyinosinic:polycytidylic acid.

    Article Snippet: Cultures were stimulated for 24 hours for mRNA extraction or for 36 hours for protein collection with the following agents: 20 μg/ml polyinosinic:polycytidylic acid (InvivoGen, San Diego, CA, USA), 100 ng/ml lipopolysaccharide from Escherichia coli (Sigma-Aldrich), 300 ng/ml bacterial lipoprotein (InvivoGen), 10 ng/ml recombinant interleukin-1β (IL-1β) (Invitrogen, Carlsbad, CA, USA), 10 ng/ml tumor necrosis factor α (TNFα) (Invitrogen) and 10 ng/ml transforming factor β (TGFβ) (Invitrogen).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Western Blot, Polymerase Chain Reaction

    Mindin mRNA expression is upregulated by CpG-ODN stimulation. About 1 × 10 5 RAW 264.7 cells were cultured in 12-well plates and stimulated with cytokines of mouse recombinant tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), toll-like receptor (TLR) ligands of Pam3-CSK4, peptidoglycans (PGN), lipopolysaccharide (LPS), flagellin and CpG-ODN 1585 (B) for 8 h, then cells were harvested for RNA isolation and relative mRNA expression of mindin was analyzed using quantitative real time polymerase chain reaction (RT-PCR). In each group, bars represent mean ± SD; b P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Mindin is upregulated during colitis and may activate NF-?B in a TLR-9 mediated manner

    doi: 10.3748/wjg.v16.i9.1070

    Figure Lengend Snippet: Mindin mRNA expression is upregulated by CpG-ODN stimulation. About 1 × 10 5 RAW 264.7 cells were cultured in 12-well plates and stimulated with cytokines of mouse recombinant tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), toll-like receptor (TLR) ligands of Pam3-CSK4, peptidoglycans (PGN), lipopolysaccharide (LPS), flagellin and CpG-ODN 1585 (B) for 8 h, then cells were harvested for RNA isolation and relative mRNA expression of mindin was analyzed using quantitative real time polymerase chain reaction (RT-PCR). In each group, bars represent mean ± SD; b P

    Article Snippet: About 1 × 105 of RAW 264.7 and CMT93 cells were cultured in 12-well plates and stimulated with the cytokines; 10 ng/mL of mouse recombinant tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) (both from R & D Systems); and different TLR ligands consisting of 0.05 μg/mL of Pam3-CSK4, 5 μg/mL of peptidoglycans (PGN), 0.1 μg/mL of LPS, 0.5 μg/mL of flagellin ( S. typhimurium ) and 5 μg/mL of CpG-ODN 1585 (all of the ligands were purchased from Invivogen), for 8 h, then cells were harvested for RNA isolation.

    Techniques: Expressing, Cell Culture, Recombinant, Isolation, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

    Concentrations of biomarkers for adults with DS with and without clinical dementia. a Aβ 40 , b Aβ 42 , c Aβ 42 /Aβ 40 , d Aβ 42 /t-tau, e t-tau, f IL1β, g IL10, h IL6, i TNFα, and j NfL. Lines indicate median value (pg/ml)

    Journal: Alzheimer's Research & Therapy

    Article Title: Plasma biomarkers for amyloid, tau, and cytokines in Down syndrome and sporadic Alzheimer’s disease

    doi: 10.1186/s13195-019-0477-0

    Figure Lengend Snippet: Concentrations of biomarkers for adults with DS with and without clinical dementia. a Aβ 40 , b Aβ 42 , c Aβ 42 /Aβ 40 , d Aβ 42 /t-tau, e t-tau, f IL1β, g IL10, h IL6, i TNFα, and j NfL. Lines indicate median value (pg/ml)

    Article Snippet: For all three groups, plasma concentrations of Aβ40 , Aβ42 , and t-tau (Human Neurology 3-Plex A assay (N3PA)), IL1β (Human IL-1β 2.0), and IL10, IL6, and TNFα (Human Cytokine 3-Plex A) were measured in duplicate using ultrasensitive Simoa immunoassays (Quanterix, Lexington, MA, USA) according to the manufacturer’s instructions at the Institute of Psychiatry, Psychology and Neuroscience, King’s College London.

    Techniques:

    Relationships between biomarkers for adults with DS. a Log Aβ 42 and log t-tau, b log t-tau and log IL1β, and c log IL1β and log IL10

    Journal: Alzheimer's Research & Therapy

    Article Title: Plasma biomarkers for amyloid, tau, and cytokines in Down syndrome and sporadic Alzheimer’s disease

    doi: 10.1186/s13195-019-0477-0

    Figure Lengend Snippet: Relationships between biomarkers for adults with DS. a Log Aβ 42 and log t-tau, b log t-tau and log IL1β, and c log IL1β and log IL10

    Article Snippet: For all three groups, plasma concentrations of Aβ40 , Aβ42 , and t-tau (Human Neurology 3-Plex A assay (N3PA)), IL1β (Human IL-1β 2.0), and IL10, IL6, and TNFα (Human Cytokine 3-Plex A) were measured in duplicate using ultrasensitive Simoa immunoassays (Quanterix, Lexington, MA, USA) according to the manufacturer’s instructions at the Institute of Psychiatry, Psychology and Neuroscience, King’s College London.

    Techniques:

    Concentrations of biomarkers for each group. a Aβ 40 , b Aβ 42 , c Aβ 42 /Aβ 40 , d Aβ 42 /t-tau, e t-tau, f IL1β, g IL10, h IL6, and i TNFα. Lines indicate median value (pg/ml)

    Journal: Alzheimer's Research & Therapy

    Article Title: Plasma biomarkers for amyloid, tau, and cytokines in Down syndrome and sporadic Alzheimer’s disease

    doi: 10.1186/s13195-019-0477-0

    Figure Lengend Snippet: Concentrations of biomarkers for each group. a Aβ 40 , b Aβ 42 , c Aβ 42 /Aβ 40 , d Aβ 42 /t-tau, e t-tau, f IL1β, g IL10, h IL6, and i TNFα. Lines indicate median value (pg/ml)

    Article Snippet: For all three groups, plasma concentrations of Aβ40 , Aβ42 , and t-tau (Human Neurology 3-Plex A assay (N3PA)), IL1β (Human IL-1β 2.0), and IL10, IL6, and TNFα (Human Cytokine 3-Plex A) were measured in duplicate using ultrasensitive Simoa immunoassays (Quanterix, Lexington, MA, USA) according to the manufacturer’s instructions at the Institute of Psychiatry, Psychology and Neuroscience, King’s College London.

    Techniques:

    The mRNA abundance of interleukin-1β (IL-1β ), tumor necrosis factor-α (TNF-α ), IL-8 and xanthine oxidoreductase ( XOR ) in udder of rats. The mRNA abundances of IL-1β , TNF-α , IL-8 and XOR were determined by RT-PCR with mammary tissues obtained after saline infusion at day 14 of gestation from pregnant rats ( n = 7) and age-matched non-pregnant rats ( n = 11). Values are presented as means ± SE. Differences between pregnant and non-pregnant rats were indicated by asterisks (**** P

    Journal: Animal Nutrition

    Article Title: Reproductive stage associated changes in plasma fatty acid profile and proinflammatory cytokine expression in rat mammary glands

    doi: 10.1016/j.aninu.2016.03.008

    Figure Lengend Snippet: The mRNA abundance of interleukin-1β (IL-1β ), tumor necrosis factor-α (TNF-α ), IL-8 and xanthine oxidoreductase ( XOR ) in udder of rats. The mRNA abundances of IL-1β , TNF-α , IL-8 and XOR were determined by RT-PCR with mammary tissues obtained after saline infusion at day 14 of gestation from pregnant rats ( n = 7) and age-matched non-pregnant rats ( n = 11). Values are presented as means ± SE. Differences between pregnant and non-pregnant rats were indicated by asterisks (**** P

    Article Snippet: 2.7 Immunohistochemistry Polyclonal antibodies combined with the avidin-biotinperoxidase complex technique were used for the immunohistochemical detection of interleukin-1β (IL-1β) (Abnova, Walnut, CA, USA) and tumor necrosis factor-α (TNF-α) (Novus, SaintCharles, MO, USA).

    Techniques: Reverse Transcription Polymerase Chain Reaction

    Immunohistochemical localization of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in mammary glands of the rats at different reproductive stages. Mammary glands were collected after saline infusion from non-pregnant rats that designated as rats at day 1 of gestation ( n = 6), pregnant rats that was infused by saline at day 14 of gestation ( n = 5) and day 3 postpartum ( n = 5). (A) Interleukin-1β and TNF-α production is presented as the average percentage of the positively stained areas. The microphotograph from one rat with the positive primary IL-1β or TNF-α antibody was visualized with diaminobenzidine reaction. The area positive for (B) IL-1β and (C) TNF-α in mammary tissue of rats at different reproductive stages was quantified by Easy Image 3000 software at a magnification of 400×. Values are presented as means ± SE. Different letters signify statistical difference ( P

    Journal: Animal Nutrition

    Article Title: Reproductive stage associated changes in plasma fatty acid profile and proinflammatory cytokine expression in rat mammary glands

    doi: 10.1016/j.aninu.2016.03.008

    Figure Lengend Snippet: Immunohistochemical localization of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in mammary glands of the rats at different reproductive stages. Mammary glands were collected after saline infusion from non-pregnant rats that designated as rats at day 1 of gestation ( n = 6), pregnant rats that was infused by saline at day 14 of gestation ( n = 5) and day 3 postpartum ( n = 5). (A) Interleukin-1β and TNF-α production is presented as the average percentage of the positively stained areas. The microphotograph from one rat with the positive primary IL-1β or TNF-α antibody was visualized with diaminobenzidine reaction. The area positive for (B) IL-1β and (C) TNF-α in mammary tissue of rats at different reproductive stages was quantified by Easy Image 3000 software at a magnification of 400×. Values are presented as means ± SE. Different letters signify statistical difference ( P

    Article Snippet: 2.7 Immunohistochemistry Polyclonal antibodies combined with the avidin-biotinperoxidase complex technique were used for the immunohistochemical detection of interleukin-1β (IL-1β) (Abnova, Walnut, CA, USA) and tumor necrosis factor-α (TNF-α) (Novus, SaintCharles, MO, USA).

    Techniques: Immunohistochemistry, Staining, Software

    Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and IL‐1β‐induced increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p

    Journal: JBMR Plus

    Article Title: A Novel Distal Enhancer Mediates Inflammation‐, PTH‐, and Early Onset Murine Kidney Disease‐Induced Expression of the Mouse Fgf23 Gene

    doi: 10.1002/jbm4.10023

    Figure Lengend Snippet: Lack of the −16kb enhancer of Fgf23 blunts the Tnfα‐ and IL‐1β‐induced increase in Fgf23 levels. Eight‐week‐old wild‐type (WT) and Fgf23 −16KO female mice were injected with 2 μg of recombinant Tnfα or vehicle, while their male littermates were injected with 50 ng/g of IL‐1β or PBS. The tissues were collected 3 hours after TNFα and 6 hours after IL‐1β injection. The Fgf23 mRNA levels were measured by RT‐PCR and represented here as the mean ± SD ( n = 5 to 9 mice/group). Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p

    Article Snippet: Reagents The following reagents were used for in vivo injections: LPS (Sigma, St. Louis, MO, USA), IL‐1β (Cell Signaling Technology, Danvers, MA, USA), and TNFα (R & D Systems, Minneapolis, MN, USA).

    Techniques: Mouse Assay, Injection, Recombinant, Reverse Transcription Polymerase Chain Reaction

    Inflammation increases Fgf23 mRNA expression in bone and non‐osseous tissues. Twelve‐month‐old female mice were injected intraperitoneally (ip) with 10 mg/kg of LPS or PBS. Eight‐week‐old female mice were injected with 2 μg of recombinant TNFα, 50 ng/g of IL‐1β, or PBS. The tissues and blood were collected 3 hours after TNFα and 6 hours after LPS or IL‐1β injection. ( A–H ) Fgf23 mRNA levels were measured by quantitative RT‐PCR and normalized to beta‐actin mRNA levels. ( I ) Blood was collected by cardiac puncture, and circulating intact FGF23 (iFGF23) levels were measured in EDTA plasma. ( A–I ) All values represent mean ± SD of 3 to 4 mice/group. All statistical comparisons were performed using Student's t test. * p

    Journal: JBMR Plus

    Article Title: A Novel Distal Enhancer Mediates Inflammation‐, PTH‐, and Early Onset Murine Kidney Disease‐Induced Expression of the Mouse Fgf23 Gene

    doi: 10.1002/jbm4.10023

    Figure Lengend Snippet: Inflammation increases Fgf23 mRNA expression in bone and non‐osseous tissues. Twelve‐month‐old female mice were injected intraperitoneally (ip) with 10 mg/kg of LPS or PBS. Eight‐week‐old female mice were injected with 2 μg of recombinant TNFα, 50 ng/g of IL‐1β, or PBS. The tissues and blood were collected 3 hours after TNFα and 6 hours after LPS or IL‐1β injection. ( A–H ) Fgf23 mRNA levels were measured by quantitative RT‐PCR and normalized to beta‐actin mRNA levels. ( I ) Blood was collected by cardiac puncture, and circulating intact FGF23 (iFGF23) levels were measured in EDTA plasma. ( A–I ) All values represent mean ± SD of 3 to 4 mice/group. All statistical comparisons were performed using Student's t test. * p

    Article Snippet: Reagents The following reagents were used for in vivo injections: LPS (Sigma, St. Louis, MO, USA), IL‐1β (Cell Signaling Technology, Danvers, MA, USA), and TNFα (R & D Systems, Minneapolis, MN, USA).

    Techniques: Expressing, Mouse Assay, Injection, Recombinant, Quantitative RT-PCR

    The −16kb enhancer of Fgf23 mediates the inflammation‐induced increase in circulating intact FGF23 levels. Eight‐ to 12‐week‐old wild‐type (WT) and Fgf23 −16KO mice were injected with 10 mg/kg of LPS (female mice) ( A ), 2 μg of recombinant TNFα (female mice) ( B ), 50 ng/g of IL‐1β (male mice) ( C ), or vehicle. Cardiac blood was collected at time of death, which was 6 hours after LPS and IL‐1β injection and 3 hours after TNFα injection. Circulating intact FGF23 (iFGF23) levels were measured by ELISA. The bars represent the mean ± SD of 4 to 9 mice/group. Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p

    Journal: JBMR Plus

    Article Title: A Novel Distal Enhancer Mediates Inflammation‐, PTH‐, and Early Onset Murine Kidney Disease‐Induced Expression of the Mouse Fgf23 Gene

    doi: 10.1002/jbm4.10023

    Figure Lengend Snippet: The −16kb enhancer of Fgf23 mediates the inflammation‐induced increase in circulating intact FGF23 levels. Eight‐ to 12‐week‐old wild‐type (WT) and Fgf23 −16KO mice were injected with 10 mg/kg of LPS (female mice) ( A ), 2 μg of recombinant TNFα (female mice) ( B ), 50 ng/g of IL‐1β (male mice) ( C ), or vehicle. Cardiac blood was collected at time of death, which was 6 hours after LPS and IL‐1β injection and 3 hours after TNFα injection. Circulating intact FGF23 (iFGF23) levels were measured by ELISA. The bars represent the mean ± SD of 4 to 9 mice/group. Statistical comparisons were performed using two‐way ANOVA with multiple comparison test using the Benjamini‐Hochberg procedure. * p

    Article Snippet: Reagents The following reagents were used for in vivo injections: LPS (Sigma, St. Louis, MO, USA), IL‐1β (Cell Signaling Technology, Danvers, MA, USA), and TNFα (R & D Systems, Minneapolis, MN, USA).

    Techniques: Mouse Assay, Injection, Recombinant, Enzyme-linked Immunosorbent Assay

    APOL1 risk variants induce inflammatory cell death (pyroptosis) in cells ( a ) Representative Western blot analysis of caspase3 (Casp3) and cleaved caspase3 (cCasp3) following transient transfection with TRE-APOL1-G0/G1/G2. UV exposure served as apoptosis positive control. n = 5. ( b ) Representative Western blot analysis of caspase1 (Casp1) and cleaved caspase1 (cCasp1) following transient transfection of TRE-APOL1-G0/G1/G2. GFP served as an APOL1 expression reference, α-tubulin as loading control and LPS as pyroptosis positive control. Western blot analysis of mature IL1β in medium from the same transfected HeLa cells. ( c ) Cell toxicity (measured by propidium iodide staining) in stably transfected TRE-GFP/APOL1-G1 HEK293 cells, with (APOL1) or without (CTL) doxycycline and in the presence of the indicated caspase1 inhibitors (concentration see Methods). Experiments were done in triplicates. Data are presented as means ± s.e.m, and Student’s t -test, P = 0.0015 (Ac-YVAD-CHO), 0.00018 (VX765) compared to APOL1. ( d ) Cell toxicity (measured by LDH release to the medium) in stably transfected TRE-GFP/APOL1-G1 HEK293 cells with (APOL1) or without (CTL) doxycycline and with indicated inhibitors (concentration see Methods). A representative experiment out of three is presented; each experiment was done in triplicates. All data are presented as means ± s.e.m, and Student’s t -test, P

    Journal: Nature medicine

    Article Title: Transgenic Expression of Human APOL1 Risk Variants in Podocytes Induces Kidney Disease in Mice

    doi: 10.1038/nm.4287

    Figure Lengend Snippet: APOL1 risk variants induce inflammatory cell death (pyroptosis) in cells ( a ) Representative Western blot analysis of caspase3 (Casp3) and cleaved caspase3 (cCasp3) following transient transfection with TRE-APOL1-G0/G1/G2. UV exposure served as apoptosis positive control. n = 5. ( b ) Representative Western blot analysis of caspase1 (Casp1) and cleaved caspase1 (cCasp1) following transient transfection of TRE-APOL1-G0/G1/G2. GFP served as an APOL1 expression reference, α-tubulin as loading control and LPS as pyroptosis positive control. Western blot analysis of mature IL1β in medium from the same transfected HeLa cells. ( c ) Cell toxicity (measured by propidium iodide staining) in stably transfected TRE-GFP/APOL1-G1 HEK293 cells, with (APOL1) or without (CTL) doxycycline and in the presence of the indicated caspase1 inhibitors (concentration see Methods). Experiments were done in triplicates. Data are presented as means ± s.e.m, and Student’s t -test, P = 0.0015 (Ac-YVAD-CHO), 0.00018 (VX765) compared to APOL1. ( d ) Cell toxicity (measured by LDH release to the medium) in stably transfected TRE-GFP/APOL1-G1 HEK293 cells with (APOL1) or without (CTL) doxycycline and with indicated inhibitors (concentration see Methods). A representative experiment out of three is presented; each experiment was done in triplicates. All data are presented as means ± s.e.m, and Student’s t -test, P

    Article Snippet: The following primary antibodies were used: GFP (immunohistochemistry (IHC), Clontech #632380, 1:500; western blot (WB), 1:3000), WT1 (IHC, Santa Cruz #M3561, 1:40), EEA1 (immunofluorescence (IF), BD transduction #610456, 1:100), Vamp8 (IF, Sigma #SAB1409943, 1:100), Rab7 (IF, Sigma #R8779, 1:100), Rab11 (IF, BD transduction #610656 1:100), LAMP2 (IF, BD transduction #555803, 1:100), LC3II (WB, Sigma #L7543, 1:1000; IF, cell signaling #27755, 1:100), STX17 (IF, Sigma #HPA001204, 1:100), APOL1 (WB, Sigma #HPA018885, 1:1000; IF, Sigma, 1:100), GM130 (IF, BD transduction #610822, 1:100), calnexin (IF, BD transduction #610523, 1:200), perilipin2 (IF, Sigma #611042, 1:500), Nephrin (IF, Fitzgerald Industries Intl Inc #20R-NP002, 1:100), cleaved-Caspase-3 (WB and IHC, cell signaling #9664, 1:1000), Caspase-1 (WB, Santa Cruz #SC515, 1:1000; IHC, Thermo Fisher #PA5-38099, 1:100), IL1β (WB, R & D #AF-201-NA, 1:1000; IHC, cell signaling #12242S, 1:100), NLRP3 (IHC, Novus #NBP2-12446, 1:100), α-tubulin (WB, Sigma #T6199, 1:3000).

    Techniques: Western Blot, Transfection, Positive Control, Expressing, Staining, Stable Transfection, CTL Assay, Concentration Assay

    Mice with podocyte specific APOL1 risk allele expression show increased podocyte loss, autophagy block and increased inflammatory cell death ( a ) Representative immunohistochemistry images of WT1 (upper), nephrin (middle) and TUNEL staining (lower) in NPHS1-rtTA/TRE-APOL1-G0/G1/G2 mouse kidney sections. ( n > 5 per line) Scale bars, 10μm. ( b ) Left panel: Quantification of stained cells per glomerulus of WT1 (a podocyte marker), in transgenic mice. n = 11 analyzed images per condition. Student’s t- test, P =0.0036 compared to NPHS1-rtTA/TRE-APOL1-G0 mice and single transgenic control mice. Right panel: Quantification of stained cells per glomerulus of TUNEL stain from transgenic mice. n = 12 analyzed images per condition. Student’s t -test, P = 0.0011 compared to NPHS1-rtTA/TRE-APOL1-G0 mice and single transgenic control. ( c ) Representative transmission EM images of podocytes from transgenic mice showing increased numbers of MVB and autophagosomes. Inserts show multivesicular bodies (*) and amphisome-like structures (**). Scale bars, 500nm. ( d ) Quantification of autophagosomes (AP) and autolysosomes (AL) in transgenic mice. Note increased ratio of AP to AL in G1 and G2 mice compared to G0 mice. n > 50 analyzed organelles per each condition. Data are presented as means ± s.e.m. and Student’s t -test, P = 8.1816e-07 (left panel), 0.037 (right panel) compared to NPHS1-rtTA/TRE-APOL1-G0 mice. ( e ) Representative immunohistochemistry images of LC3II from transgenic mice. Note the increased podocyte stain in G1 and G2 mice compared to G0 mice, suggesting increase in autophagic vacuole content. ( n > 5 per line) Scale bars, 10μm. ( f–h ) Representative immunohistochemistry images of IL1β, NLRP3 and caspase1, showing increased stain of pyroptosis proteins in podocytes of G1 and G2 mice, compared to G0 mice. ( n > 5 per line) Scale bars, 10 μm. ( j ) Representative immunohistochemistry images of cleaved caspase3 from transgenic mice. Note lack of detectable stain, indicating apoptosis is not significantly induced in transgenic mice. ( n > 5 per line) Scale bars, 10 μm. ( i ) Differential expression of a set of genes by RNA-seq analysis in kidneys of NPHS1-rtTA/TRE-APOL1-G0/G1/G2 mice. Single transgenic littermates and NPHS1-rtTA/TRE-APOL1-G0 are controls.

    Journal: Nature medicine

    Article Title: Transgenic Expression of Human APOL1 Risk Variants in Podocytes Induces Kidney Disease in Mice

    doi: 10.1038/nm.4287

    Figure Lengend Snippet: Mice with podocyte specific APOL1 risk allele expression show increased podocyte loss, autophagy block and increased inflammatory cell death ( a ) Representative immunohistochemistry images of WT1 (upper), nephrin (middle) and TUNEL staining (lower) in NPHS1-rtTA/TRE-APOL1-G0/G1/G2 mouse kidney sections. ( n > 5 per line) Scale bars, 10μm. ( b ) Left panel: Quantification of stained cells per glomerulus of WT1 (a podocyte marker), in transgenic mice. n = 11 analyzed images per condition. Student’s t- test, P =0.0036 compared to NPHS1-rtTA/TRE-APOL1-G0 mice and single transgenic control mice. Right panel: Quantification of stained cells per glomerulus of TUNEL stain from transgenic mice. n = 12 analyzed images per condition. Student’s t -test, P = 0.0011 compared to NPHS1-rtTA/TRE-APOL1-G0 mice and single transgenic control. ( c ) Representative transmission EM images of podocytes from transgenic mice showing increased numbers of MVB and autophagosomes. Inserts show multivesicular bodies (*) and amphisome-like structures (**). Scale bars, 500nm. ( d ) Quantification of autophagosomes (AP) and autolysosomes (AL) in transgenic mice. Note increased ratio of AP to AL in G1 and G2 mice compared to G0 mice. n > 50 analyzed organelles per each condition. Data are presented as means ± s.e.m. and Student’s t -test, P = 8.1816e-07 (left panel), 0.037 (right panel) compared to NPHS1-rtTA/TRE-APOL1-G0 mice. ( e ) Representative immunohistochemistry images of LC3II from transgenic mice. Note the increased podocyte stain in G1 and G2 mice compared to G0 mice, suggesting increase in autophagic vacuole content. ( n > 5 per line) Scale bars, 10μm. ( f–h ) Representative immunohistochemistry images of IL1β, NLRP3 and caspase1, showing increased stain of pyroptosis proteins in podocytes of G1 and G2 mice, compared to G0 mice. ( n > 5 per line) Scale bars, 10 μm. ( j ) Representative immunohistochemistry images of cleaved caspase3 from transgenic mice. Note lack of detectable stain, indicating apoptosis is not significantly induced in transgenic mice. ( n > 5 per line) Scale bars, 10 μm. ( i ) Differential expression of a set of genes by RNA-seq analysis in kidneys of NPHS1-rtTA/TRE-APOL1-G0/G1/G2 mice. Single transgenic littermates and NPHS1-rtTA/TRE-APOL1-G0 are controls.

    Article Snippet: The following primary antibodies were used: GFP (immunohistochemistry (IHC), Clontech #632380, 1:500; western blot (WB), 1:3000), WT1 (IHC, Santa Cruz #M3561, 1:40), EEA1 (immunofluorescence (IF), BD transduction #610456, 1:100), Vamp8 (IF, Sigma #SAB1409943, 1:100), Rab7 (IF, Sigma #R8779, 1:100), Rab11 (IF, BD transduction #610656 1:100), LAMP2 (IF, BD transduction #555803, 1:100), LC3II (WB, Sigma #L7543, 1:1000; IF, cell signaling #27755, 1:100), STX17 (IF, Sigma #HPA001204, 1:100), APOL1 (WB, Sigma #HPA018885, 1:1000; IF, Sigma, 1:100), GM130 (IF, BD transduction #610822, 1:100), calnexin (IF, BD transduction #610523, 1:200), perilipin2 (IF, Sigma #611042, 1:500), Nephrin (IF, Fitzgerald Industries Intl Inc #20R-NP002, 1:100), cleaved-Caspase-3 (WB and IHC, cell signaling #9664, 1:1000), Caspase-1 (WB, Santa Cruz #SC515, 1:1000; IHC, Thermo Fisher #PA5-38099, 1:100), IL1β (WB, R & D #AF-201-NA, 1:1000; IHC, cell signaling #12242S, 1:100), NLRP3 (IHC, Novus #NBP2-12446, 1:100), α-tubulin (WB, Sigma #T6199, 1:3000).

    Techniques: Mouse Assay, Expressing, Blocking Assay, Immunohistochemistry, TUNEL Assay, Staining, Marker, Transgenic Assay, Transmission Assay, RNA Sequencing Assay

    Cytosolic adaptor caspase recruitment domain 9 (CARD9) deficiency inhibits angiotensin II (Ang II) infusion–induced proinflammatory cell infiltration and expression of inflammatory cytokines. ( a ) Heart sections from wild-type (WT) and CARD9 −/− mice treated with saline or Ang II at 1,500 ng/kg/min for 7 days were stained with hematoxylin and eosin (HE). ( b – d ) Heart sections from WT and CARD9 −/− mice were detected by immunohistochemistry with antibodies against Mac-2, interleukin-1β (IL-1β), and connective tissue growth factor (CTGF) (top). Bar graph shows quantified areas (bottom). Bar = 50 µm. Data represent mean ± s.e.m. ( n = 5). * P

    Journal: American Journal of Hypertension

    Article Title: Proinflammatory Protein CARD9 Is Essential for Infiltration of Monocytic Fibroblast Precursors and Cardiac Fibrosis Caused by Angiotensin II Infusion

    doi: 10.1038/ajh.2011.42

    Figure Lengend Snippet: Cytosolic adaptor caspase recruitment domain 9 (CARD9) deficiency inhibits angiotensin II (Ang II) infusion–induced proinflammatory cell infiltration and expression of inflammatory cytokines. ( a ) Heart sections from wild-type (WT) and CARD9 −/− mice treated with saline or Ang II at 1,500 ng/kg/min for 7 days were stained with hematoxylin and eosin (HE). ( b – d ) Heart sections from WT and CARD9 −/− mice were detected by immunohistochemistry with antibodies against Mac-2, interleukin-1β (IL-1β), and connective tissue growth factor (CTGF) (top). Bar graph shows quantified areas (bottom). Bar = 50 µm. Data represent mean ± s.e.m. ( n = 5). * P

    Article Snippet: Immunohistochemical analysis involved antibodies against CARD9 (1:200 dilution), Mac-2 (1:400 dilution), interleukin-1β (IL-1β) (1:200 dilution), connective tissue growth factor (CTGF) (1:200 dilution) and transforming growth factor-β (TGF-β) (1:300 dilution; all Santa Cruz Biotechnology, Santa Cruz, CA); and collagen I (1:1,000 dilution).

    Techniques: Expressing, Mouse Assay, Staining, Immunohistochemistry

    Catabolic regulation of LRP5 mediates via β-catenin-Tcf/Lef signaling. (A) Chondrocytes were transfected with 1 μg of empty vector (EV) or pSPORT6- Lrp5 plus the TOPflash or FOPflash reporter constructs. After 24 hours, transcriptional activation by β-catenin was determined by luciferase reporter gene assays ( n = 7 independent experiments). (B) Chondrocytes obtained from wild-type (WT) and Lrp5 -/- mice were treated with 1 ng/ml interleukin 1β (IL-1β), 50 ng/ml Wnt3a or 500 ng/ml Wnt7a, and transcriptional activation by β-catenin was evaluated by luciferase reporter gene assays ( n = 6). Values are expressed as means ± SEM (* P

    Journal: Arthritis Research & Therapy

    Article Title: Low-density lipoprotein receptor-related protein 5 governs Wnt-mediated osteoarthritic cartilage destruction

    doi: 10.1186/ar4466

    Figure Lengend Snippet: Catabolic regulation of LRP5 mediates via β-catenin-Tcf/Lef signaling. (A) Chondrocytes were transfected with 1 μg of empty vector (EV) or pSPORT6- Lrp5 plus the TOPflash or FOPflash reporter constructs. After 24 hours, transcriptional activation by β-catenin was determined by luciferase reporter gene assays ( n = 7 independent experiments). (B) Chondrocytes obtained from wild-type (WT) and Lrp5 -/- mice were treated with 1 ng/ml interleukin 1β (IL-1β), 50 ng/ml Wnt3a or 500 ng/ml Wnt7a, and transcriptional activation by β-catenin was evaluated by luciferase reporter gene assays ( n = 6). Values are expressed as means ± SEM (* P

    Article Snippet: On culture day 3, the cells were treated with recombinant interleukin 1β (IL-1β) (Calbiochem, San Diego, CA, USA), Wnt3a or Wnt7a (R & D Systems, Minneapolis, MN, USA) for 24 hours.

    Techniques: Transfection, Plasmid Preparation, Construct, Activation Assay, Luciferase, Mouse Assay

    Upregulation of LRP5 in interleukin 1β-treated mouse articular chondrocytes. (A) Mesenchymal cells from mouse embryos were maintained as micromass cultures for 12 days, and the expression levels of Col2a1 , Mmp13 , Lrp5 , Lrp6 and Gapdh were detected by RT-PCR. LRP5 expression was confirmed by Western blot analysis. (B) and (C) Primary cultured mouse articular chondrocytes were treated with the indicated concentrations of interleukin 1β (IL-1β) for 24 hours (B) or with 1 ng/ml IL-1β for the indicated periods (C), and RT-PCR, quantitative RT-PCR and Western blot analysis were carried out. Values are presented as means ± SEM (* P

    Journal: Arthritis Research & Therapy

    Article Title: Low-density lipoprotein receptor-related protein 5 governs Wnt-mediated osteoarthritic cartilage destruction

    doi: 10.1186/ar4466

    Figure Lengend Snippet: Upregulation of LRP5 in interleukin 1β-treated mouse articular chondrocytes. (A) Mesenchymal cells from mouse embryos were maintained as micromass cultures for 12 days, and the expression levels of Col2a1 , Mmp13 , Lrp5 , Lrp6 and Gapdh were detected by RT-PCR. LRP5 expression was confirmed by Western blot analysis. (B) and (C) Primary cultured mouse articular chondrocytes were treated with the indicated concentrations of interleukin 1β (IL-1β) for 24 hours (B) or with 1 ng/ml IL-1β for the indicated periods (C), and RT-PCR, quantitative RT-PCR and Western blot analysis were carried out. Values are presented as means ± SEM (* P

    Article Snippet: On culture day 3, the cells were treated with recombinant interleukin 1β (IL-1β) (Calbiochem, San Diego, CA, USA), Wnt3a or Wnt7a (R & D Systems, Minneapolis, MN, USA) for 24 hours.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Cell Culture, Quantitative RT-PCR

    PmpG-1-vaults activate the NLRP3 inflammasome and induce IL-1β secretion, as measured by an ELISA assay

    Journal: Vaccine

    Article Title: Activation of the NLRP3 inflammasome by vault nanoparticles expressing a chlamydial epitope

    doi: 10.1016/j.vaccine.2014.11.028

    Figure Lengend Snippet: PmpG-1-vaults activate the NLRP3 inflammasome and induce IL-1β secretion, as measured by an ELISA assay

    Article Snippet: To detect mature IL-1β, the blot was probed with IL-1β antibody (Cell Signaling) at a 1:1000 dilution, and then incubated again with 1:10000 dilution of anti-mouse secondary antibody (Santa Cruz Biotechnology).

    Techniques: Enzyme-linked Immunosorbent Assay