monosodium urate msu crystals  (InvivoGen)

 
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    Name:
    MSU Crystals
    Description:
    Monosodium Urate Crystals
    Catalog Number:
    tlrl-msu-25
    Price:
    None
    Size:
    25 mg
    Category:
    MSU Crystals Inflammasome Inducers PRR Ligands
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    Structured Review

    InvivoGen monosodium urate msu crystals
    iNKT Cells Induce IL-1β Secretion by <t>BMDCs</t> in a CD1d-Dependent Manner (A and B) BMDCs were left unprimed or primed with Pam3CSK4 (P3C; 0.5 μg/mL) for 24 h and subsequently co-cultured with iNKT cells for 24 h in the (A) absence or (B) presence of αGalCer (50 ng/mL). IL-1β release was quantified by ELISA. As a control, BMDCs were primed with P3C for 4 h, followed by 6-h stimulation with <t>MSU</t> (100 μg/mL). The same concentrations of P3C, αGalCer, and MSU were used throughout all experiments. (C and D) P3C-primed BMDCs were cultured alone or with iNKT cells for the indicated times in the presence or absence of αGalCer. IL-1β release was quantified by ELISA (C), and cell-associated and extracellular IL-1β were analyzed by immunoblot (D). As a control, BMDCs were primed with P3C for 4 h, followed by 1-h stimulation with nigericin (10 μM). (E) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with iNKT cells for the indicated times in the absence or presence of αGalCer. IL-1β release was quantified by ELISA. P3C-primed BMDCs stimulated with nigericin were used as a control. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments. *p
    Monosodium Urate Crystals
    https://www.bioz.com/result/monosodium urate msu crystals/product/InvivoGen
    Average 99 stars, based on 1 article reviews
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    monosodium urate msu crystals - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "A Two-Cell Model for IL-1β Release Mediated by Death-Receptor Signaling"

    Article Title: A Two-Cell Model for IL-1β Release Mediated by Death-Receptor Signaling

    Journal: Cell reports

    doi: 10.1016/j.celrep.2020.03.030

    iNKT Cells Induce IL-1β Secretion by BMDCs in a CD1d-Dependent Manner (A and B) BMDCs were left unprimed or primed with Pam3CSK4 (P3C; 0.5 μg/mL) for 24 h and subsequently co-cultured with iNKT cells for 24 h in the (A) absence or (B) presence of αGalCer (50 ng/mL). IL-1β release was quantified by ELISA. As a control, BMDCs were primed with P3C for 4 h, followed by 6-h stimulation with MSU (100 μg/mL). The same concentrations of P3C, αGalCer, and MSU were used throughout all experiments. (C and D) P3C-primed BMDCs were cultured alone or with iNKT cells for the indicated times in the presence or absence of αGalCer. IL-1β release was quantified by ELISA (C), and cell-associated and extracellular IL-1β were analyzed by immunoblot (D). As a control, BMDCs were primed with P3C for 4 h, followed by 1-h stimulation with nigericin (10 μM). (E) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with iNKT cells for the indicated times in the absence or presence of αGalCer. IL-1β release was quantified by ELISA. P3C-primed BMDCs stimulated with nigericin were used as a control. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments. *p
    Figure Legend Snippet: iNKT Cells Induce IL-1β Secretion by BMDCs in a CD1d-Dependent Manner (A and B) BMDCs were left unprimed or primed with Pam3CSK4 (P3C; 0.5 μg/mL) for 24 h and subsequently co-cultured with iNKT cells for 24 h in the (A) absence or (B) presence of αGalCer (50 ng/mL). IL-1β release was quantified by ELISA. As a control, BMDCs were primed with P3C for 4 h, followed by 6-h stimulation with MSU (100 μg/mL). The same concentrations of P3C, αGalCer, and MSU were used throughout all experiments. (C and D) P3C-primed BMDCs were cultured alone or with iNKT cells for the indicated times in the presence or absence of αGalCer. IL-1β release was quantified by ELISA (C), and cell-associated and extracellular IL-1β were analyzed by immunoblot (D). As a control, BMDCs were primed with P3C for 4 h, followed by 1-h stimulation with nigericin (10 μM). (E) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with iNKT cells for the indicated times in the absence or presence of αGalCer. IL-1β release was quantified by ELISA. P3C-primed BMDCs stimulated with nigericin were used as a control. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments. *p

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay, Mouse Assay

    iNKT Cells Employ the Death Receptor Ligand FasL to Instruct TLR-Primed BMDCs to Release IL-1β (A) iNKT cells were cultured in the presence of increasing concentrations of plate-bound anti-CD3 for 24 h, after which their cell-free Supes were used to stimulate P3C-primed BMDCs for 24 h. IL-1β (left) and interferon γ (IFN-γ; right) in the Supes were quantified by ELISA. Nigericin-stimulated BMDCs were used as a control. (B) The ImmGen database was mined to compare the expression of cell-surface proteins (Gene Ontology term “cell surface,” GO0009986) expressed by iNKT cells versus other αβ T cells. The red dots represent genes expressed by iNKT cells with a fold change of greater than 5 compared with all other αβ T cells. Expression values comparing class means for specific T cell populations are shown in the right panel. (C) P3C-primed BMDCs were co-cultured with iNKT cells for 24 h in the presence of a FasL-blocking antibody (aFasL mAb) or an isotype control. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (D) P3C-primed BMDCs from WT and Fas −/− mice were co-cultured with iNKT cells for 24 h. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (E) Resting iNKT cells were stained with a FasL-specific antibody or an isotype control and subsequently analyzed by flow cytometry. The left panel represents surface staining, whereas the right panel represents permeabilized iNKT cells. (F) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with CellTrace Violet-labeled-iNKT cells for the indicated times in the presence or absence of αGalCer, and the expression of FasL on the surface of iNKT cells was analyzed by flow cytometry. Numbers on the right of each histogram represent mean fluorescence intensity values. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments.
    Figure Legend Snippet: iNKT Cells Employ the Death Receptor Ligand FasL to Instruct TLR-Primed BMDCs to Release IL-1β (A) iNKT cells were cultured in the presence of increasing concentrations of plate-bound anti-CD3 for 24 h, after which their cell-free Supes were used to stimulate P3C-primed BMDCs for 24 h. IL-1β (left) and interferon γ (IFN-γ; right) in the Supes were quantified by ELISA. Nigericin-stimulated BMDCs were used as a control. (B) The ImmGen database was mined to compare the expression of cell-surface proteins (Gene Ontology term “cell surface,” GO0009986) expressed by iNKT cells versus other αβ T cells. The red dots represent genes expressed by iNKT cells with a fold change of greater than 5 compared with all other αβ T cells. Expression values comparing class means for specific T cell populations are shown in the right panel. (C) P3C-primed BMDCs were co-cultured with iNKT cells for 24 h in the presence of a FasL-blocking antibody (aFasL mAb) or an isotype control. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (D) P3C-primed BMDCs from WT and Fas −/− mice were co-cultured with iNKT cells for 24 h. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (E) Resting iNKT cells were stained with a FasL-specific antibody or an isotype control and subsequently analyzed by flow cytometry. The left panel represents surface staining, whereas the right panel represents permeabilized iNKT cells. (F) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with CellTrace Violet-labeled-iNKT cells for the indicated times in the presence or absence of αGalCer, and the expression of FasL on the surface of iNKT cells was analyzed by flow cytometry. Numbers on the right of each histogram represent mean fluorescence intensity values. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments.

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Blocking Assay, Mouse Assay, Staining, Flow Cytometry, Labeling, Fluorescence

    2) Product Images from "Inflammasome-Induced IL-1β Secretion in Microglia Is Characterized by Delayed Kinetics and Is Only Partially Dependent on Inflammatory Caspases"

    Article Title: Inflammasome-Induced IL-1β Secretion in Microglia Is Characterized by Delayed Kinetics and Is Only Partially Dependent on Inflammatory Caspases

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2510-14.2015

    Kinetics of IL-1β secretion by LPS-primed microglia differ from that by LPS-primed hematopoietic macrophages. A , Silica-induced (500 μg/ml, 6 h) IL-1β secretion in LPS-primed (100 ng/ml, indicated time points) microglia and BMDMs. Data of one representative donor of ≥3 independent experiments are shown and are represented as mean ± SD. B , Analysis of the ratio of inflammasome-induced IL-1β secretion after 16 h to inflammasome-induced IL-1β secretion after 4 h. Open symbols, Silica-induced IL-1β secretion; closed symbols, MSU-induced IL-1β secretion; * p
    Figure Legend Snippet: Kinetics of IL-1β secretion by LPS-primed microglia differ from that by LPS-primed hematopoietic macrophages. A , Silica-induced (500 μg/ml, 6 h) IL-1β secretion in LPS-primed (100 ng/ml, indicated time points) microglia and BMDMs. Data of one representative donor of ≥3 independent experiments are shown and are represented as mean ± SD. B , Analysis of the ratio of inflammasome-induced IL-1β secretion after 16 h to inflammasome-induced IL-1β secretion after 4 h. Open symbols, Silica-induced IL-1β secretion; closed symbols, MSU-induced IL-1β secretion; * p

    Techniques Used:

    Related Articles

    Produced:

    Article Title: Recombinant human proteoglycan-4 reduces phagocytosis of urate crystals and downstream nuclear factor kappa B and inflammasome activation and production of cytokines and chemokines in human and murine macrophages
    Article Snippet: .. THP-1 macrophages were treated with endotoxin-free MSU crystals (100μg/ml; Invivogen, USA) ± bovine submaxillary mucin (BSM; molecular mass > 1000 KDa) (Sigma Aldrich) (25 μg/ml) or rhPRG4 (molecular mass is approximately 240 KDa) (100 μg/ml) for 2 and 4 h at 37 °C. rhPRG4 is an endotoxin-free full-length product produced by CHO-M cells (Lubris, Framingham, MA, USA) [ ]. ..

    Marker:

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR
    Article Snippet: .. Both MSU crystals and IL-1β significantly induced cell death in cytotrophoblast cultures, as seen by the 3.3-fold and 2.7-fold increase, respectively, in the percentage of cells positive for the apoptotic marker M30 at 48h ( ). .. Treatment of MSU crystals-exposed cytotrophoblasts with IL-1Ra was protective, with decreased percentage of M30+ apoptotic cells ( ).

    Injection:

    Article Title: The Immunomodulatory Metabolite Itaconate Modifies NLRP3 and Inhibits Inflammasome Activation
    Article Snippet: .. MSU-Induced Peritonitis Model 6-week old female C57BL/6J mice were injected intraperitoneally with a mixture of 4-OI (50 mg/kg) in 60% cyclodextrin in PBS and MSU crystals (30mg/kg, Invivogen) suspended in PBS for 6 h. Mice were euthanized in a CO2 chamber and peritoneal lavage was performed using 2.5 mL PBS. .. The cells in the lavage fluid were pelleted and the supernatant was removed and analyzed by ELISA for IL-1β and IL-6 concentration.

    other:

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR
    Article Snippet: Since IL-1β is a well-known mediator of MSU crystals actions in immune cells, and it was strongly induced in term cytotrophoblasts in response to MSU crystals, we addressed the mechanisms of IL-1β production.

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR
    Article Snippet: Consistent with these findings in cytotrophoblast, treatment with MSU crystals or IL-1β induced apoptosis in term placental explants, ( ) which was mainly observed in cytotrophoblast cells (arrowheads in ).

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR
    Article Snippet: This was predominantly observed within the junctional zone, where all doses of MSU crystals led to a significant elevation of monocytes/macrophages (CD68+ cells) and within the labyrinth (fetal side), where only the highest dose of MSU crystals induced a significant increased in CD68+ monocytes/macrophages ( ).

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis
    Article Snippet: MSU crystals can induce a variety of cytokines, including IL-1β, IL-6, IL-8, and TNF-α [ ].

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR
    Article Snippet: The secretion of IL-1β induced by MSU crystals in cytotrophoblasts was dependent on the common processor of pro-IL-1β, caspase-1, which was ascertained using a caspase-1 inhibitor ( ).

    Mouse Assay:

    Article Title: The Immunomodulatory Metabolite Itaconate Modifies NLRP3 and Inhibits Inflammasome Activation
    Article Snippet: .. MSU-Induced Peritonitis Model 6-week old female C57BL/6J mice were injected intraperitoneally with a mixture of 4-OI (50 mg/kg) in 60% cyclodextrin in PBS and MSU crystals (30mg/kg, Invivogen) suspended in PBS for 6 h. Mice were euthanized in a CO2 chamber and peritoneal lavage was performed using 2.5 mL PBS. .. The cells in the lavage fluid were pelleted and the supernatant was removed and analyzed by ELISA for IL-1β and IL-6 concentration.

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    InvivoGen monosodium urate crystal msu
    Effects of GW-A2 on NLRP3 inflammasome activation. ( A and B ) J774A.1 macrophages were incubated for 30 min with or without 2 μM of GW-A2, for 5.5 h with or without 0.1 μg/ml of E . coli LPS, and then for 30 min with or without 5 mM of <t>ATP</t> ( A ) or 10 μM of nigericin ( B ). The levels of IL-1β in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. ( C — F ) J774A.1 macrophages were incubated for 5.5 h with 0.1 μg/ml of E . coli LPS, for 30 min with or without 2 μM of GW-A2, and then for 30 min with or without 5 mM of ATP ( C and E ), 10 μM of nigericin ( D ) or 100 μg/ml of <t>MSU</t> ( F ). The levels of IL-1β and TNF-α in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. The data are expressed as the mean ± SD of three independent experiments. The Western blotting results shown are a representative experiment of three independent experiments. *, ** and *** indicate significant differences, representing p
    Monosodium Urate Crystal Msu, supplied by InvivoGen, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monosodium urate crystal msu/product/InvivoGen
    Average 99 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    monosodium urate crystal msu - by Bioz Stars, 2020-09
    99/100 stars
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    Effects of GW-A2 on NLRP3 inflammasome activation. ( A and B ) J774A.1 macrophages were incubated for 30 min with or without 2 μM of GW-A2, for 5.5 h with or without 0.1 μg/ml of E . coli LPS, and then for 30 min with or without 5 mM of ATP ( A ) or 10 μM of nigericin ( B ). The levels of IL-1β in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. ( C — F ) J774A.1 macrophages were incubated for 5.5 h with 0.1 μg/ml of E . coli LPS, for 30 min with or without 2 μM of GW-A2, and then for 30 min with or without 5 mM of ATP ( C and E ), 10 μM of nigericin ( D ) or 100 μg/ml of MSU ( F ). The levels of IL-1β and TNF-α in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. The data are expressed as the mean ± SD of three independent experiments. The Western blotting results shown are a representative experiment of three independent experiments. *, ** and *** indicate significant differences, representing p

    Journal: PLoS ONE

    Article Title: A synthetic cationic antimicrobial peptide inhibits inflammatory response and the NLRP3 inflammasome by neutralizing LPS and ATP

    doi: 10.1371/journal.pone.0182057

    Figure Lengend Snippet: Effects of GW-A2 on NLRP3 inflammasome activation. ( A and B ) J774A.1 macrophages were incubated for 30 min with or without 2 μM of GW-A2, for 5.5 h with or without 0.1 μg/ml of E . coli LPS, and then for 30 min with or without 5 mM of ATP ( A ) or 10 μM of nigericin ( B ). The levels of IL-1β in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. ( C — F ) J774A.1 macrophages were incubated for 5.5 h with 0.1 μg/ml of E . coli LPS, for 30 min with or without 2 μM of GW-A2, and then for 30 min with or without 5 mM of ATP ( C and E ), 10 μM of nigericin ( D ) or 100 μg/ml of MSU ( F ). The levels of IL-1β and TNF-α in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. The data are expressed as the mean ± SD of three independent experiments. The Western blotting results shown are a representative experiment of three independent experiments. *, ** and *** indicate significant differences, representing p

    Article Snippet: Pam3CSK4 (tlrl-pms), ATP (tlrl-atp), nigericin (tlrl-nig) and monosodium urate crystal (MSU) (tlrl-msu) were purchased from InvivoGen (San Diego, CA, USA).

    Techniques: Activation Assay, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot

    Effect of APO on cell viability and IL-1 β secretion in LPS-primed BMDMs. (a) BMDMs were treated with the indicated concentration of APO for 24 h. Cell viability was measured by the LDH assay. (b–f) LPS-primed BMDMs were pretreated with APO or zVAD (20 μ M), and IL-1 β secretion was determined by ELISA upon stimulation with (b) 150 μ g/mL silica for 3 h, (c) 10 μ M nigericin for 1 h, (d) 5 mM ATP for 1 h, (e) 2 μ g/mL poly (dA:dT) for 1 h, and (f) 1.5 μ g/mL flagellin for 3 h. (g) TNF- α release from LPS-primed BMDMs was determined upon stimulation with 10 μ M nigericin for 1 h. (h) The impact of APO on MSU-induced IL-1 β production in mice. Data were expressed as the mean ± SEM ( n = 3). Statistical analysis was performed using Student's t -test. ∗ p

    Journal: Mediators of Inflammation

    Article Title: Artemisia Extract Suppresses NLRP3 and AIM2 Inflammasome Activation by Inhibition of ASC Phosphorylation

    doi: 10.1155/2018/6054069

    Figure Lengend Snippet: Effect of APO on cell viability and IL-1 β secretion in LPS-primed BMDMs. (a) BMDMs were treated with the indicated concentration of APO for 24 h. Cell viability was measured by the LDH assay. (b–f) LPS-primed BMDMs were pretreated with APO or zVAD (20 μ M), and IL-1 β secretion was determined by ELISA upon stimulation with (b) 150 μ g/mL silica for 3 h, (c) 10 μ M nigericin for 1 h, (d) 5 mM ATP for 1 h, (e) 2 μ g/mL poly (dA:dT) for 1 h, and (f) 1.5 μ g/mL flagellin for 3 h. (g) TNF- α release from LPS-primed BMDMs was determined upon stimulation with 10 μ M nigericin for 1 h. (h) The impact of APO on MSU-induced IL-1 β production in mice. Data were expressed as the mean ± SEM ( n = 3). Statistical analysis was performed using Student's t -test. ∗ p

    Article Snippet: Silica crystals (tlrl-sio), nigericin (tlrl-nig), ultrapure flagellin from Salmonella typhimurium (tlrl-epstfla), poly(deoxyadenylic-deoxythymidylic) acid (poly dA:dT, tlrl-patn), monosodium urate (MSU) crystal (tlrl-msu), and carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (zVAD-FMK, tlrl-vad) were purchased from InvivoGen (San Diego, CA, USA).

    Techniques: Concentration Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay

    iNKT Cells Induce IL-1β Secretion by BMDCs in a CD1d-Dependent Manner (A and B) BMDCs were left unprimed or primed with Pam3CSK4 (P3C; 0.5 μg/mL) for 24 h and subsequently co-cultured with iNKT cells for 24 h in the (A) absence or (B) presence of αGalCer (50 ng/mL). IL-1β release was quantified by ELISA. As a control, BMDCs were primed with P3C for 4 h, followed by 6-h stimulation with MSU (100 μg/mL). The same concentrations of P3C, αGalCer, and MSU were used throughout all experiments. (C and D) P3C-primed BMDCs were cultured alone or with iNKT cells for the indicated times in the presence or absence of αGalCer. IL-1β release was quantified by ELISA (C), and cell-associated and extracellular IL-1β were analyzed by immunoblot (D). As a control, BMDCs were primed with P3C for 4 h, followed by 1-h stimulation with nigericin (10 μM). (E) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with iNKT cells for the indicated times in the absence or presence of αGalCer. IL-1β release was quantified by ELISA. P3C-primed BMDCs stimulated with nigericin were used as a control. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments. *p

    Journal: Cell reports

    Article Title: A Two-Cell Model for IL-1β Release Mediated by Death-Receptor Signaling

    doi: 10.1016/j.celrep.2020.03.030

    Figure Lengend Snippet: iNKT Cells Induce IL-1β Secretion by BMDCs in a CD1d-Dependent Manner (A and B) BMDCs were left unprimed or primed with Pam3CSK4 (P3C; 0.5 μg/mL) for 24 h and subsequently co-cultured with iNKT cells for 24 h in the (A) absence or (B) presence of αGalCer (50 ng/mL). IL-1β release was quantified by ELISA. As a control, BMDCs were primed with P3C for 4 h, followed by 6-h stimulation with MSU (100 μg/mL). The same concentrations of P3C, αGalCer, and MSU were used throughout all experiments. (C and D) P3C-primed BMDCs were cultured alone or with iNKT cells for the indicated times in the presence or absence of αGalCer. IL-1β release was quantified by ELISA (C), and cell-associated and extracellular IL-1β were analyzed by immunoblot (D). As a control, BMDCs were primed with P3C for 4 h, followed by 1-h stimulation with nigericin (10 μM). (E) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with iNKT cells for the indicated times in the absence or presence of αGalCer. IL-1β release was quantified by ELISA. P3C-primed BMDCs stimulated with nigericin were used as a control. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments. *p

    Article Snippet: For inflammasome activation controls, BMDCs were primed for 4 hours and treated with monosodium urate (MSU) crystals (Invivogen, 100 μg/ml) for 6 hours or nigericin (Invivogen, 10 μM) for 1 hour.

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Mouse Assay

    iNKT Cells Employ the Death Receptor Ligand FasL to Instruct TLR-Primed BMDCs to Release IL-1β (A) iNKT cells were cultured in the presence of increasing concentrations of plate-bound anti-CD3 for 24 h, after which their cell-free Supes were used to stimulate P3C-primed BMDCs for 24 h. IL-1β (left) and interferon γ (IFN-γ; right) in the Supes were quantified by ELISA. Nigericin-stimulated BMDCs were used as a control. (B) The ImmGen database was mined to compare the expression of cell-surface proteins (Gene Ontology term “cell surface,” GO0009986) expressed by iNKT cells versus other αβ T cells. The red dots represent genes expressed by iNKT cells with a fold change of greater than 5 compared with all other αβ T cells. Expression values comparing class means for specific T cell populations are shown in the right panel. (C) P3C-primed BMDCs were co-cultured with iNKT cells for 24 h in the presence of a FasL-blocking antibody (aFasL mAb) or an isotype control. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (D) P3C-primed BMDCs from WT and Fas −/− mice were co-cultured with iNKT cells for 24 h. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (E) Resting iNKT cells were stained with a FasL-specific antibody or an isotype control and subsequently analyzed by flow cytometry. The left panel represents surface staining, whereas the right panel represents permeabilized iNKT cells. (F) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with CellTrace Violet-labeled-iNKT cells for the indicated times in the presence or absence of αGalCer, and the expression of FasL on the surface of iNKT cells was analyzed by flow cytometry. Numbers on the right of each histogram represent mean fluorescence intensity values. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments.

    Journal: Cell reports

    Article Title: A Two-Cell Model for IL-1β Release Mediated by Death-Receptor Signaling

    doi: 10.1016/j.celrep.2020.03.030

    Figure Lengend Snippet: iNKT Cells Employ the Death Receptor Ligand FasL to Instruct TLR-Primed BMDCs to Release IL-1β (A) iNKT cells were cultured in the presence of increasing concentrations of plate-bound anti-CD3 for 24 h, after which their cell-free Supes were used to stimulate P3C-primed BMDCs for 24 h. IL-1β (left) and interferon γ (IFN-γ; right) in the Supes were quantified by ELISA. Nigericin-stimulated BMDCs were used as a control. (B) The ImmGen database was mined to compare the expression of cell-surface proteins (Gene Ontology term “cell surface,” GO0009986) expressed by iNKT cells versus other αβ T cells. The red dots represent genes expressed by iNKT cells with a fold change of greater than 5 compared with all other αβ T cells. Expression values comparing class means for specific T cell populations are shown in the right panel. (C) P3C-primed BMDCs were co-cultured with iNKT cells for 24 h in the presence of a FasL-blocking antibody (aFasL mAb) or an isotype control. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (D) P3C-primed BMDCs from WT and Fas −/− mice were co-cultured with iNKT cells for 24 h. IL-1β release was quantified by ELISA. MSU-treated BMDCs were used as a control. (E) Resting iNKT cells were stained with a FasL-specific antibody or an isotype control and subsequently analyzed by flow cytometry. The left panel represents surface staining, whereas the right panel represents permeabilized iNKT cells. (F) P3C-primed BMDCs from WT and CD1d −/− mice were co-cultured with CellTrace Violet-labeled-iNKT cells for the indicated times in the presence or absence of αGalCer, and the expression of FasL on the surface of iNKT cells was analyzed by flow cytometry. Numbers on the right of each histogram represent mean fluorescence intensity values. Error bars represent mean + SD of triplicate samples. Each panel is representative of at least three different experiments.

    Article Snippet: For inflammasome activation controls, BMDCs were primed for 4 hours and treated with monosodium urate (MSU) crystals (Invivogen, 100 μg/ml) for 6 hours or nigericin (Invivogen, 10 μM) for 1 hour.

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Blocking Assay, Mouse Assay, Staining, Flow Cytometry, Labeling, Fluorescence

    Effect of APO on cell viability and IL-1 β secretion in LPS-primed BMDMs. (a) BMDMs were treated with the indicated concentration of APO for 24 h. Cell viability was measured by the LDH assay. (b–f) LPS-primed BMDMs were pretreated with APO or zVAD (20 μ M), and IL-1 β secretion was determined by ELISA upon stimulation with (b) 150 μ g/mL silica for 3 h, (c) 10 μ M nigericin for 1 h, (d) 5 mM ATP for 1 h, (e) 2 μ g/mL poly (dA:dT) for 1 h, and (f) 1.5 μ g/mL flagellin for 3 h. (g) TNF- α release from LPS-primed BMDMs was determined upon stimulation with 10 μ M nigericin for 1 h. (h) The impact of APO on MSU-induced IL-1 β production in mice. Data were expressed as the mean ± SEM ( n = 3). Statistical analysis was performed using Student's t -test. ∗ p

    Journal: Mediators of Inflammation

    Article Title: Artemisia Extract Suppresses NLRP3 and AIM2 Inflammasome Activation by Inhibition of ASC Phosphorylation

    doi: 10.1155/2018/6054069

    Figure Lengend Snippet: Effect of APO on cell viability and IL-1 β secretion in LPS-primed BMDMs. (a) BMDMs were treated with the indicated concentration of APO for 24 h. Cell viability was measured by the LDH assay. (b–f) LPS-primed BMDMs were pretreated with APO or zVAD (20 μ M), and IL-1 β secretion was determined by ELISA upon stimulation with (b) 150 μ g/mL silica for 3 h, (c) 10 μ M nigericin for 1 h, (d) 5 mM ATP for 1 h, (e) 2 μ g/mL poly (dA:dT) for 1 h, and (f) 1.5 μ g/mL flagellin for 3 h. (g) TNF- α release from LPS-primed BMDMs was determined upon stimulation with 10 μ M nigericin for 1 h. (h) The impact of APO on MSU-induced IL-1 β production in mice. Data were expressed as the mean ± SEM ( n = 3). Statistical analysis was performed using Student's t -test. ∗ p

    Article Snippet: Silica crystals (tlrl-sio), nigericin (tlrl-nig), ultrapure flagellin from Salmonella typhimurium (tlrl-epstfla), poly(deoxyadenylic-deoxythymidylic) acid (poly dA:dT, tlrl-patn), monosodium urate (MSU) crystal (tlrl-msu), and carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (zVAD-FMK, tlrl-vad) were purchased from InvivoGen (San Diego, CA, USA).

    Techniques: Concentration Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay