msu crystals  (InvivoGen)

 
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  • 99
    Name:
    MSU Crystals
    Description:
    Monosodium Urate Crystals
    Catalog Number:
    tlrl-msu-25
    Price:
    None
    Size:
    25 mg
    Category:
    MSU Crystals Inflammasome Inducers PRR Ligands
    Buy from Supplier


    Structured Review

    InvivoGen msu crystals
    <t>MSU</t> triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), <t>PMA</t> (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Monosodium Urate Crystals
    https://www.bioz.com/result/msu crystals/product/InvivoGen
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    msu crystals - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals"

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21113750

    MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used: Amplification, Produced

    MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.
    Figure Legend Snippet: MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.

    Techniques Used: Fluorescence

    MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used:

    2) Product Images from "The Immunomodulatory Metabolite Itaconate Modifies NLRP3 and Inhibits Inflammasome Activation"

    Article Title: The Immunomodulatory Metabolite Itaconate Modifies NLRP3 and Inhibits Inflammasome Activation

    Journal: Cell Metabolism

    doi: 10.1016/j.cmet.2020.07.016

    4-OI Reduces Inflammation in a Murine In Vivo Model of Peritonitis and Blocks NLRP3 Inflammasome Activation in Healthy Human and CAPS PBMCs (A–C) IL-1β concentration (A), IL-6 concentration (B), and neutrophil number (C) in the peritoneal lavage fluid of mice injected for 6 h with MSU crystals (30 mg/kg) ± 4-OI (50 mg/kg) (n = 3 for PBS groups, n = 8 for MSU groups). (D) LPS or Pam3CSK4 (14 h) and nigericin (2 h) induced IL-1β release (n = 5 for LPS + nigericin, n = 3 for Pam3CSK4 + nigericin) ± 4-OI or 4-O-2-MS (both 250 μM) from healthy human PBMCs. (E and F) Immunoblot analysis (E) and quantification by densitometry (F, n = 3) of pro- and mature IL-1β protein in lysates and supernatants of human PBMCs treated with LPS (14 h) and nigericin (2 h) ± 4-OI (250 μM). (G) LPS (1 h) induced IL-1β release (n = 3) ± 4-OI (250 μM) or MCC950 (500 nM) from PBMCs isolated from CAPS patients. ∗ p
    Figure Legend Snippet: 4-OI Reduces Inflammation in a Murine In Vivo Model of Peritonitis and Blocks NLRP3 Inflammasome Activation in Healthy Human and CAPS PBMCs (A–C) IL-1β concentration (A), IL-6 concentration (B), and neutrophil number (C) in the peritoneal lavage fluid of mice injected for 6 h with MSU crystals (30 mg/kg) ± 4-OI (50 mg/kg) (n = 3 for PBS groups, n = 8 for MSU groups). (D) LPS or Pam3CSK4 (14 h) and nigericin (2 h) induced IL-1β release (n = 5 for LPS + nigericin, n = 3 for Pam3CSK4 + nigericin) ± 4-OI or 4-O-2-MS (both 250 μM) from healthy human PBMCs. (E and F) Immunoblot analysis (E) and quantification by densitometry (F, n = 3) of pro- and mature IL-1β protein in lysates and supernatants of human PBMCs treated with LPS (14 h) and nigericin (2 h) ± 4-OI (250 μM). (G) LPS (1 h) induced IL-1β release (n = 3) ± 4-OI (250 μM) or MCC950 (500 nM) from PBMCs isolated from CAPS patients. ∗ p

    Techniques Used: In Vivo, Activation Assay, Concentration Assay, Mouse Assay, Injection, Isolation

    3) Product Images from "MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis"

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-017-1418-6

    MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P
    Figure Legend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P

    Techniques Used: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P
    Figure Legend Snippet: Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P

    Techniques Used: Expressing, Concentration Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    4) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    5) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    6) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    7) Product Images from "In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals"

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21113750

    MSU-induced ROS originate from the NADPH-oxidase. ( A ) Neutrophils pre-treated with the NADPH-oxidase inhibitors DPI (10 µg/mL) or GSK (20 µg/mL) did not produce icROS in response to MSU crystals (300 μg/mL); peak values +/− SD are shown ( n = 3). ( B ) Neutrophils from a patient with CGD (broken line) did not produce icROS in response to MSU crystals (300 µg/mL) as compared to control neutrophils (solid line). Arrows indicate addition of stimuli.
    Figure Legend Snippet: MSU-induced ROS originate from the NADPH-oxidase. ( A ) Neutrophils pre-treated with the NADPH-oxidase inhibitors DPI (10 µg/mL) or GSK (20 µg/mL) did not produce icROS in response to MSU crystals (300 μg/mL); peak values +/− SD are shown ( n = 3). ( B ) Neutrophils from a patient with CGD (broken line) did not produce icROS in response to MSU crystals (300 µg/mL) as compared to control neutrophils (solid line). Arrows indicate addition of stimuli.

    Techniques Used:

    MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used:

    8) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    9) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of CD68+ macrophage in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of CD68+ macrophage in the placenta following MSU crystals treatment.

    Techniques Used:

    10) Product Images from "In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals"

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21113750

    MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.
    Figure Legend Snippet: MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.

    Techniques Used: Fluorescence

    MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used:

    In vivo transmigrated neutrophils are not primed for NET formation. Sytox green fluorescence assay was used to evaluate NET formation of tissue neutrophils from skin chamber fluid ( A ) or synovial fluid ( B ) after stimulation with MSU crystals (300 µg/mL). Tissue neutrophils (broken lines) formed NETs in a similar manner as blood neutrophils (solid lines) from the same donor assayed in parallel. Shown are representative curves for skin chamber neutrophils ( A ), and synovial fluid neutrophils ( B ), and the mean fluorescence values +/− SEM at 3 h from three independent experiments of each type of tissue neutrophils.
    Figure Legend Snippet: In vivo transmigrated neutrophils are not primed for NET formation. Sytox green fluorescence assay was used to evaluate NET formation of tissue neutrophils from skin chamber fluid ( A ) or synovial fluid ( B ) after stimulation with MSU crystals (300 µg/mL). Tissue neutrophils (broken lines) formed NETs in a similar manner as blood neutrophils (solid lines) from the same donor assayed in parallel. Shown are representative curves for skin chamber neutrophils ( A ), and synovial fluid neutrophils ( B ), and the mean fluorescence values +/− SEM at 3 h from three independent experiments of each type of tissue neutrophils.

    Techniques Used: In Vivo, Fluorescence

    11) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    12) Product Images from "In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals"

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21113750

    MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used: Amplification, Produced

    MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.
    Figure Legend Snippet: MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.

    Techniques Used: Fluorescence

    MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used:

    13) Product Images from "MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis"

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-017-1418-6

    MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P
    Figure Legend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P

    Techniques Used: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P
    Figure Legend Snippet: Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P

    Techniques Used: Expressing, Concentration Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    14) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    15) Product Images from "In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals"

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21113750

    MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used: Amplification, Produced

    MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.
    Figure Legend Snippet: MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.

    Techniques Used: Fluorescence

    MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Figure Legend Snippet: MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Techniques Used:

    16) Product Images from "MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis"

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-017-1418-6

    MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P
    Figure Legend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P

    Techniques Used: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    Effects of monosodium urate (MSU) crystals on expression of microRNAs (miRNAs, miR) in monocytic THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The expression levels of miRNAs were detected by quantitative real-time polymerase chain reaction( a - e ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P
    Figure Legend Snippet: Effects of monosodium urate (MSU) crystals on expression of microRNAs (miRNAs, miR) in monocytic THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The expression levels of miRNAs were detected by quantitative real-time polymerase chain reaction( a - e ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P

    Techniques Used: Expressing, Concentration Assay

    MicroRNA (miR, miRNA)-488 and miR-920 inhibit monosodium urate (MSU)-induced interleukin (IL)-1β protein expression in monocytic THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of IL-1β. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a ) or Western blotting ( c ). The cell culture supernatants were collected to detect the concentrations of IL-1β by enzyme-linked immunosorbent assay ( b ). d Densitometric analysis of immunoblot band intensities for IL-1β normalized by β-actin. Values are expressed as mean ± SEM of three independent experiments. # P
    Figure Legend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 inhibit monosodium urate (MSU)-induced interleukin (IL)-1β protein expression in monocytic THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of IL-1β. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a ) or Western blotting ( c ). The cell culture supernatants were collected to detect the concentrations of IL-1β by enzyme-linked immunosorbent assay ( b ). d Densitometric analysis of immunoblot band intensities for IL-1β normalized by β-actin. Values are expressed as mean ± SEM of three independent experiments. # P

    Techniques Used: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

    Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P
    Figure Legend Snippet: Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P

    Techniques Used: Expressing, Concentration Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    17) Product Images from "Insight into Neutrophil Extracellular Traps through Systematic Evaluation of Citrullination and Peptidylarginine Deiminases"

    Article Title: Insight into Neutrophil Extracellular Traps through Systematic Evaluation of Citrullination and Peptidylarginine Deiminases

    Journal: Journal of Immunology Research

    doi: 10.1155/2019/2160192

    PAD4 is required for the formation of citrullinated NETs in murine neutrophils. Bone marrow neutrophils from PAD4 +/+ and PAD4 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD4 +/+ and PAD4 −/− mice. For all panels: n = 4; ∗ p
    Figure Legend Snippet: PAD4 is required for the formation of citrullinated NETs in murine neutrophils. Bone marrow neutrophils from PAD4 +/+ and PAD4 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD4 +/+ and PAD4 −/− mice. For all panels: n = 4; ∗ p

    Techniques Used: Mouse Assay, Staining

    PAD2 is not required for the formation of NETs in murine neutrophils. Bone marrow neutrophils from PAD2 +/+ and PAD2 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD2 +/+ and PAD2 −/− mice. For all panels: n = 4; no comparisons were significant.
    Figure Legend Snippet: PAD2 is not required for the formation of NETs in murine neutrophils. Bone marrow neutrophils from PAD2 +/+ and PAD2 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD2 +/+ and PAD2 −/− mice. For all panels: n = 4; no comparisons were significant.

    Techniques Used: Mouse Assay, Staining

    Induction of NETs in human neutrophils. Human neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, or C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with C. albicans and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 9; ∗ p
    Figure Legend Snippet: Induction of NETs in human neutrophils. Human neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, or C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with C. albicans and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 9; ∗ p

    Techniques Used: Staining, Labeling, Negative Control

    Induction of NETs in murine neutrophils. Murine neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with ionomycin and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. Enlarged insets demonstrate donut-like structures (doNETs). The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 6; ∗ p
    Figure Legend Snippet: Induction of NETs in murine neutrophils. Murine neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with ionomycin and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. Enlarged insets demonstrate donut-like structures (doNETs). The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 6; ∗ p

    Techniques Used: Staining, Labeling, Negative Control

    18) Product Images from "Insight into Neutrophil Extracellular Traps through Systematic Evaluation of Citrullination and Peptidylarginine Deiminases"

    Article Title: Insight into Neutrophil Extracellular Traps through Systematic Evaluation of Citrullination and Peptidylarginine Deiminases

    Journal: Journal of Immunology Research

    doi: 10.1155/2019/2160192

    PAD4 is required for the formation of citrullinated NETs in murine neutrophils. Bone marrow neutrophils from PAD4 +/+ and PAD4 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD4 +/+ and PAD4 −/− mice. For all panels: n = 4; ∗ p
    Figure Legend Snippet: PAD4 is required for the formation of citrullinated NETs in murine neutrophils. Bone marrow neutrophils from PAD4 +/+ and PAD4 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD4 +/+ and PAD4 −/− mice. For all panels: n = 4; ∗ p

    Techniques Used: Mouse Assay, Staining

    PAD2 is not required for the formation of NETs in murine neutrophils. Bone marrow neutrophils from PAD2 +/+ and PAD2 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD2 +/+ and PAD2 −/− mice. For all panels: n = 4; no comparisons were significant.
    Figure Legend Snippet: PAD2 is not required for the formation of NETs in murine neutrophils. Bone marrow neutrophils from PAD2 +/+ and PAD2 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD2 +/+ and PAD2 −/− mice. For all panels: n = 4; no comparisons were significant.

    Techniques Used: Mouse Assay, Staining

    Induction of NETs in human neutrophils. Human neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, or C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with C. albicans and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 9; ∗ p
    Figure Legend Snippet: Induction of NETs in human neutrophils. Human neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, or C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with C. albicans and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 9; ∗ p

    Techniques Used: Staining, Labeling, Negative Control

    Induction of NETs in murine neutrophils. Murine neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with ionomycin and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. Enlarged insets demonstrate donut-like structures (doNETs). The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 6; ∗ p
    Figure Legend Snippet: Induction of NETs in murine neutrophils. Murine neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with ionomycin and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. Enlarged insets demonstrate donut-like structures (doNETs). The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 6; ∗ p

    Techniques Used: Staining, Labeling, Negative Control

    19) Product Images from "Insight into Neutrophil Extracellular Traps through Systematic Evaluation of Citrullination and Peptidylarginine Deiminases"

    Article Title: Insight into Neutrophil Extracellular Traps through Systematic Evaluation of Citrullination and Peptidylarginine Deiminases

    Journal: Journal of Immunology Research

    doi: 10.1155/2019/2160192

    PAD4 is required for the formation of citrullinated NETs in murine neutrophils. Bone marrow neutrophils from PAD4 +/+ and PAD4 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD4 +/+ and PAD4 −/− mice. For all panels: n = 4; ∗ p
    Figure Legend Snippet: PAD4 is required for the formation of citrullinated NETs in murine neutrophils. Bone marrow neutrophils from PAD4 +/+ and PAD4 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD4 +/+ and PAD4 −/− mice. For all panels: n = 4; ∗ p

    Techniques Used: Mouse Assay, Staining

    PAD2 is not required for the formation of NETs in murine neutrophils. Bone marrow neutrophils from PAD2 +/+ and PAD2 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD2 +/+ and PAD2 −/− mice. For all panels: n = 4; no comparisons were significant.
    Figure Legend Snippet: PAD2 is not required for the formation of NETs in murine neutrophils. Bone marrow neutrophils from PAD2 +/+ and PAD2 −/− mice were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared between PAD2 +/+ and PAD2 −/− mice. For all panels: n = 4; no comparisons were significant.

    Techniques Used: Mouse Assay, Staining

    Induction of NETs in human neutrophils. Human neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, or C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with C. albicans and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 9; ∗ p
    Figure Legend Snippet: Induction of NETs in human neutrophils. Human neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, or C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with C. albicans and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 9; ∗ p

    Techniques Used: Staining, Labeling, Negative Control

    Induction of NETs in murine neutrophils. Murine neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with ionomycin and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. Enlarged insets demonstrate donut-like structures (doNETs). The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 6; ∗ p
    Figure Legend Snippet: Induction of NETs in murine neutrophils. Murine neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with ionomycin and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, scale bar = 50 μ M. Enlarged insets demonstrate donut-like structures (doNETs). The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: n = 6; ∗ p

    Techniques Used: Staining, Labeling, Negative Control

    20) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    21) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    22) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    23) Product Images from "Acidic preconditioning of endothelial colony-forming cells (ECFC) promote vasculogenesis under proinflammatory and high glucose conditions in vitro and in vivo"

    Article Title: Acidic preconditioning of endothelial colony-forming cells (ECFC) promote vasculogenesis under proinflammatory and high glucose conditions in vitro and in vivo

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-0872-7

    Monosodium urate (MSU) induced endothelial colony-forming cell (EFCF) necrosis. a ECFC were treated with MSU at the indicated concentration and the cell death was analyzed by fluoresce microscopy after 24 h ( n = 5). b Kinetics of ECFC death were studied by fluorescence microscopy ( n = 4). c Representative images of viable (V) and necrotic (N) cells are shown. Original magnification, 1200×. Scale bar = 10 μm. d The percentage of cell death was measured by fluorescence microscopy in the presence or absence of Z-VAD-fmk (30 μM). The percentage of positive cells for active caspase-3 ( e ), interleukin (IL)-1β levels ( f ), and reactive oxygen species (ROS) formation ( g ) were analyzed by flow cytometry after 24 h ( n = 3). * p
    Figure Legend Snippet: Monosodium urate (MSU) induced endothelial colony-forming cell (EFCF) necrosis. a ECFC were treated with MSU at the indicated concentration and the cell death was analyzed by fluoresce microscopy after 24 h ( n = 5). b Kinetics of ECFC death were studied by fluorescence microscopy ( n = 4). c Representative images of viable (V) and necrotic (N) cells are shown. Original magnification, 1200×. Scale bar = 10 μm. d The percentage of cell death was measured by fluorescence microscopy in the presence or absence of Z-VAD-fmk (30 μM). The percentage of positive cells for active caspase-3 ( e ), interleukin (IL)-1β levels ( f ), and reactive oxygen species (ROS) formation ( g ) were analyzed by flow cytometry after 24 h ( n = 3). * p

    Techniques Used: Concentration Assay, Microscopy, Fluorescence, Flow Cytometry, Cytometry

    24) Product Images from "MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis"

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-017-1418-6

    MicroRNA (miR, miRNA)-488 and miR-920 inhibit monosodium urate (MSU)-induced interleukin (IL)-1β protein expression in monocytic THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of IL-1β. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a ) or Western blotting ( c ). The cell culture supernatants were collected to detect the concentrations of IL-1β by enzyme-linked immunosorbent assay ( b ). d Densitometric analysis of immunoblot band intensities for IL-1β normalized by β-actin. Values are expressed as mean ± SEM of three independent experiments. # P
    Figure Legend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 inhibit monosodium urate (MSU)-induced interleukin (IL)-1β protein expression in monocytic THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of IL-1β. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a ) or Western blotting ( c ). The cell culture supernatants were collected to detect the concentrations of IL-1β by enzyme-linked immunosorbent assay ( b ). d Densitometric analysis of immunoblot band intensities for IL-1β normalized by β-actin. Values are expressed as mean ± SEM of three independent experiments. # P

    Techniques Used: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

    Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P
    Figure Legend Snippet: Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P

    Techniques Used: Expressing, Concentration Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    25) Product Images from "CD44 Receptor Mediates Urate Crystal Phagocytosis by Macrophages and Regulates Inflammation in A Murine Peritoneal Model of Acute Gout"

    Article Title: CD44 Receptor Mediates Urate Crystal Phagocytosis by Macrophages and Regulates Inflammation in A Murine Peritoneal Model of Acute Gout

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-62727-z

    Impact of anti-CD44 antibody treatment on NLRP3 inflammasome activation and nuclear factor kappa B (NFκB) p65 subunit nuclear translocation in differentiated human THP-1 macrophages following incubation with monosodium urate monohydrate (MSU) crystals and role of intracellular protein-phosphatase 2A (PP2A) in mediating anti-CD44 antibody’s effect. We utilized the IM7 clone, which recognizes all CD44 isoforms and causes enzyme-mediated shedding of the extracellular domain. THP-1 macrophages were treated with MSU crystals (100 μg/mL) ± anti-CD44 or isotype control (IC) antibodies (2 μg/mL for both antibodies). NF-κB p65 nuclear levels were determined following a 1-hour incubation using a DNA binding assay. Pro-caspase-1 and the p10 subunit of active caspase-1 were determined using Western Blotting. Active caspase-1 activity was determined using a specific activity assay. Cellular pro-IL-1β and mature IL-1β levels were determined using an ELISA. PP2A inhibition was performed using okadiac acid (5 nM). Data from 3 independent experiments are presented with mean and S.D. highlighted. *p
    Figure Legend Snippet: Impact of anti-CD44 antibody treatment on NLRP3 inflammasome activation and nuclear factor kappa B (NFκB) p65 subunit nuclear translocation in differentiated human THP-1 macrophages following incubation with monosodium urate monohydrate (MSU) crystals and role of intracellular protein-phosphatase 2A (PP2A) in mediating anti-CD44 antibody’s effect. We utilized the IM7 clone, which recognizes all CD44 isoforms and causes enzyme-mediated shedding of the extracellular domain. THP-1 macrophages were treated with MSU crystals (100 μg/mL) ± anti-CD44 or isotype control (IC) antibodies (2 μg/mL for both antibodies). NF-κB p65 nuclear levels were determined following a 1-hour incubation using a DNA binding assay. Pro-caspase-1 and the p10 subunit of active caspase-1 were determined using Western Blotting. Active caspase-1 activity was determined using a specific activity assay. Cellular pro-IL-1β and mature IL-1β levels were determined using an ELISA. PP2A inhibition was performed using okadiac acid (5 nM). Data from 3 independent experiments are presented with mean and S.D. highlighted. *p

    Techniques Used: Activation Assay, Translocation Assay, Incubation, DNA Binding Assay, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay, Inhibition

    Impact of monosodium urate monohydrate (MSU) crystal treatment on CD44 receptor expression and internalization in differentiated human THP-1 macrophages. CD44 expression was determined using qPCR, flow cytometry and ELISA at 3, 6, 12 and 24 hours following MSU treatment. CD44 cytosolic internalization was determined using confocal microscopy, and intracellular CD44 staining intensity was determined following incubation with MSU crystals for 3 and 6 hours. Data are from 3 independent experiments and are presented with mean and S.D. highlighted. *p
    Figure Legend Snippet: Impact of monosodium urate monohydrate (MSU) crystal treatment on CD44 receptor expression and internalization in differentiated human THP-1 macrophages. CD44 expression was determined using qPCR, flow cytometry and ELISA at 3, 6, 12 and 24 hours following MSU treatment. CD44 cytosolic internalization was determined using confocal microscopy, and intracellular CD44 staining intensity was determined following incubation with MSU crystals for 3 and 6 hours. Data are from 3 independent experiments and are presented with mean and S.D. highlighted. *p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Confocal Microscopy, Staining, Incubation

    Impact of antibody-mediated CD44 receptor shedding on monosodium urate monohydrate (MSU) crystal phagocytosis by THP-1 macrophages, expression and production of interleukin-1 beta (IL-1β) and interleukin-8 (IL8). We utilized the IM7 clone, which recognizes all CD44 isoforms and causes enzyme-mediated shedding of the extracellular domain. THP-1 macrophages were incubated with MSU crystals (100 μg/mL) ± anti-CD44 or isotype control (IC) antibodies (2 μg/mL for both antibodies). MSU phagocytosis was determined by the change in side scatter in macrophages following a 4-hour incubation and expressed as percent positive cells that internalized MSU crystals. Representative flow cytometry plots are shown in supplementary figure 2. Gene expression and production of cytokines were determined following a 6-hour incubation. Data from 3–5 independent experiments are presented with mean and S.D. highlighted. Dashed line represents gene expression in untreated controls. *p
    Figure Legend Snippet: Impact of antibody-mediated CD44 receptor shedding on monosodium urate monohydrate (MSU) crystal phagocytosis by THP-1 macrophages, expression and production of interleukin-1 beta (IL-1β) and interleukin-8 (IL8). We utilized the IM7 clone, which recognizes all CD44 isoforms and causes enzyme-mediated shedding of the extracellular domain. THP-1 macrophages were incubated with MSU crystals (100 μg/mL) ± anti-CD44 or isotype control (IC) antibodies (2 μg/mL for both antibodies). MSU phagocytosis was determined by the change in side scatter in macrophages following a 4-hour incubation and expressed as percent positive cells that internalized MSU crystals. Representative flow cytometry plots are shown in supplementary figure 2. Gene expression and production of cytokines were determined following a 6-hour incubation. Data from 3–5 independent experiments are presented with mean and S.D. highlighted. Dashed line represents gene expression in untreated controls. *p

    Techniques Used: Expressing, Incubation, Flow Cytometry

    Impact of intraperitoneal administration of monosodium urate monohydrate (MSU) crystals on inflammatory cell infiltration and production of interleukin-1 beta (IL-1β) and interleukin-1 receptor antagonist (IL-1Ra) in wildtype, Cd44 −/− and Nlrp3 −/− mice. MSU (2 mg in 200 μl PBS) or PBS (200 μl) were administered via the intraperitoneal route in wildtype (n = 8), Cd44 −/− (n = 8) or Nlrp3 −/− (n = 6) and peritoneal lavaging was performed at 4 hours. Total peritoneal lavage cell counts were determined. The number of infiltrated neutrophils and monocytes were determined using flow cytometry and probing for neutrophil markers; Ly6B.2 and Ly6G and monocyte markers; Cd11b and Ly6C. Representative flow cytometry plots of neutrophil markers are shown in supplementary figure 3. Peritoneal lavage IL-1β and IL-1Ra levels were determined by ELISA and the IL-1Ra/IL-1β ratios were computed. *p
    Figure Legend Snippet: Impact of intraperitoneal administration of monosodium urate monohydrate (MSU) crystals on inflammatory cell infiltration and production of interleukin-1 beta (IL-1β) and interleukin-1 receptor antagonist (IL-1Ra) in wildtype, Cd44 −/− and Nlrp3 −/− mice. MSU (2 mg in 200 μl PBS) or PBS (200 μl) were administered via the intraperitoneal route in wildtype (n = 8), Cd44 −/− (n = 8) or Nlrp3 −/− (n = 6) and peritoneal lavaging was performed at 4 hours. Total peritoneal lavage cell counts were determined. The number of infiltrated neutrophils and monocytes were determined using flow cytometry and probing for neutrophil markers; Ly6B.2 and Ly6G and monocyte markers; Cd11b and Ly6C. Representative flow cytometry plots of neutrophil markers are shown in supplementary figure 3. Peritoneal lavage IL-1β and IL-1Ra levels were determined by ELISA and the IL-1Ra/IL-1β ratios were computed. *p

    Techniques Used: Mouse Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

    Role of CD44 receptor in mediating monosodium urate monohydrate (MSU) crystal phagocytosis and downstream inflammation in bone marrow derived macrophages (BMDMs) from Cd44 +/+ and Cd44 −/− mice and impact of antibody-mediated receptor shedding on MSU phagocytosis by murine macrophages. BMDMs were treated with MSU (100 μg/mL) for 4 hours to evaluate phagocytosis and for 6 hours to evaluate interleukin-1 beta (IL-1β) gene expression and production. BMDMs were stimulated with lipopolysaccharide (LPS; 100 ng/ml) for 72 hours as a positive control. Lactate dehydrogenase (LDH) release from BMDMs was determined at 1 and 4 hours following incubation with MSU crystals and LDH activity level was used to calculate percent cytotoxicity. We utilized the IM7 clone, which recognizes all CD44 isoforms and causes shedding of the extracellular domain. Anti-CD44 and isotype control (IC) antibodies (2 μg/mL for both antibodies) were incubated with J774A.1 murine macrophages for 4 hours. Data from 3–5 independent experiments are presented with mean and S.D. highlighted. *p
    Figure Legend Snippet: Role of CD44 receptor in mediating monosodium urate monohydrate (MSU) crystal phagocytosis and downstream inflammation in bone marrow derived macrophages (BMDMs) from Cd44 +/+ and Cd44 −/− mice and impact of antibody-mediated receptor shedding on MSU phagocytosis by murine macrophages. BMDMs were treated with MSU (100 μg/mL) for 4 hours to evaluate phagocytosis and for 6 hours to evaluate interleukin-1 beta (IL-1β) gene expression and production. BMDMs were stimulated with lipopolysaccharide (LPS; 100 ng/ml) for 72 hours as a positive control. Lactate dehydrogenase (LDH) release from BMDMs was determined at 1 and 4 hours following incubation with MSU crystals and LDH activity level was used to calculate percent cytotoxicity. We utilized the IM7 clone, which recognizes all CD44 isoforms and causes shedding of the extracellular domain. Anti-CD44 and isotype control (IC) antibodies (2 μg/mL for both antibodies) were incubated with J774A.1 murine macrophages for 4 hours. Data from 3–5 independent experiments are presented with mean and S.D. highlighted. *p

    Techniques Used: Derivative Assay, Mouse Assay, Expressing, Positive Control, Incubation, Activity Assay

    Impact of anti-CD44 antibody treatment on inflammatory cell infiltration and production of interleukin-1 beta (IL-1β) in murine peritoneal monosodium urate monohydrate (MSU) crystal inflammation model. MSU crystals (2 mg in 200 μL PBS), vehicle (Veh.; 200 μL PBS), anti-CD44 and isotype control (IC) antibodies (50 μg in 200 μL PBS) were administered via the intraperitoneal route. Experimental groups included untreated controls (n = 4), MSU + Veh. (n = 4), MSU + Anti-CD44 antibody (n = 5) and MSU + IC antibody (n = 3). Peritoneal lavaging was performed at 4 hours. Total peritoneal lavage cell counts were determined. The number of infiltrated neutrophils and monocytes were determined using flow cytometry and probing for neutrophil markers; Ly6B.2 and Ly6G and monocyte markers; Cd11b and Ly6C. Representative flow cytometry plots of neutrophil and monocytes markers are shown in Supplementary Fig. 4 . Peritoneal lavage IL-1β levels were determined by ELISA. *p
    Figure Legend Snippet: Impact of anti-CD44 antibody treatment on inflammatory cell infiltration and production of interleukin-1 beta (IL-1β) in murine peritoneal monosodium urate monohydrate (MSU) crystal inflammation model. MSU crystals (2 mg in 200 μL PBS), vehicle (Veh.; 200 μL PBS), anti-CD44 and isotype control (IC) antibodies (50 μg in 200 μL PBS) were administered via the intraperitoneal route. Experimental groups included untreated controls (n = 4), MSU + Veh. (n = 4), MSU + Anti-CD44 antibody (n = 5) and MSU + IC antibody (n = 3). Peritoneal lavaging was performed at 4 hours. Total peritoneal lavage cell counts were determined. The number of infiltrated neutrophils and monocytes were determined using flow cytometry and probing for neutrophil markers; Ly6B.2 and Ly6G and monocyte markers; Cd11b and Ly6C. Representative flow cytometry plots of neutrophil and monocytes markers are shown in Supplementary Fig. 4 . Peritoneal lavage IL-1β levels were determined by ELISA. *p

    Techniques Used: Flow Cytometry, Enzyme-linked Immunosorbent Assay

    26) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of CD68+ macrophage in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of CD68+ macrophage in the placenta following MSU crystals treatment.

    Techniques Used:

    27) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    28) Product Images from "MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis"

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-017-1418-6

    MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P
    Figure Legend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P

    Techniques Used: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    MicroRNA (miR, miRNA)-488 and miR-920 inhibit monosodium urate (MSU)-induced interleukin (IL)-1β protein expression in monocytic THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of IL-1β. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a ) or Western blotting ( c ). The cell culture supernatants were collected to detect the concentrations of IL-1β by enzyme-linked immunosorbent assay ( b ). d Densitometric analysis of immunoblot band intensities for IL-1β normalized by β-actin. Values are expressed as mean ± SEM of three independent experiments. # P
    Figure Legend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 inhibit monosodium urate (MSU)-induced interleukin (IL)-1β protein expression in monocytic THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of IL-1β. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a ) or Western blotting ( c ). The cell culture supernatants were collected to detect the concentrations of IL-1β by enzyme-linked immunosorbent assay ( b ). d Densitometric analysis of immunoblot band intensities for IL-1β normalized by β-actin. Values are expressed as mean ± SEM of three independent experiments. # P

    Techniques Used: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

    29) Product Images from "Acidic preconditioning of endothelial colony-forming cells (ECFC) promote vasculogenesis under proinflammatory and high glucose conditions in vitro and in vivo"

    Article Title: Acidic preconditioning of endothelial colony-forming cells (ECFC) promote vasculogenesis under proinflammatory and high glucose conditions in vitro and in vivo

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-0872-7

    Monosodium urate (MSU) induced endothelial colony-forming cell (EFCF) necrosis. a ECFC were treated with MSU at the indicated concentration and the cell death was analyzed by fluoresce microscopy after 24 h ( n = 5). b Kinetics of ECFC death were studied by fluorescence microscopy ( n = 4). c Representative images of viable (V) and necrotic (N) cells are shown. Original magnification, 1200×. Scale bar = 10 μm. d The percentage of cell death was measured by fluorescence microscopy in the presence or absence of Z-VAD-fmk (30 μM). The percentage of positive cells for active caspase-3 ( e ), interleukin (IL)-1β levels ( f ), and reactive oxygen species (ROS) formation ( g ) were analyzed by flow cytometry after 24 h ( n = 3). * p
    Figure Legend Snippet: Monosodium urate (MSU) induced endothelial colony-forming cell (EFCF) necrosis. a ECFC were treated with MSU at the indicated concentration and the cell death was analyzed by fluoresce microscopy after 24 h ( n = 5). b Kinetics of ECFC death were studied by fluorescence microscopy ( n = 4). c Representative images of viable (V) and necrotic (N) cells are shown. Original magnification, 1200×. Scale bar = 10 μm. d The percentage of cell death was measured by fluorescence microscopy in the presence or absence of Z-VAD-fmk (30 μM). The percentage of positive cells for active caspase-3 ( e ), interleukin (IL)-1β levels ( f ), and reactive oxygen species (ROS) formation ( g ) were analyzed by flow cytometry after 24 h ( n = 3). * p

    Techniques Used: Concentration Assay, Microscopy, Fluorescence, Flow Cytometry, Cytometry

    30) Product Images from "Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR"

    Article Title: Uric acid crystals induces placental inflammation and alters trophoblast function via an IL-1-dependent pathway: implication for FGR

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

    doi: 10.4049/jimmunol.1601179

    Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.
    Figure Legend Snippet: Histological analysis of IL-1β staining in the placenta following MSU crystals treatment.

    Techniques Used: Staining

    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 msu crystals
    <t>MSU</t> triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), <t>PMA</t> (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.
    Msu Crystals, supplied by InvivoGen, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Journal: International Journal of Molecular Sciences

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    doi: 10.3390/ijms21113750

    Figure Lengend Snippet: MSU triggered intracellular ROS is inhibited by cytochalasin B (CytB) and colchicine (col). ( A ) Neutrophils were pretreated with cytochalsin B (5 µg/mL), an inhibitor of actin polymerization, and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and intracellular ROS production (icROS) was evaluated using luminol amplified CL. Cytochalasin B-treated cells produced lower amounts of icROS in response to MSU crystals and S. aureus as compared to untreated cells, while icROS production in response to PMA was not affected by the presence of cytochalasin B. Peak CL values are shown. ( B ) Neutrophils were pretreated with colchicine (1 µg/mL) and stimulated with either MSU crystals (300 µg/mL), PMA (50 nM) or opsonized S. aureus and icROS production was evaluated using luminol amplified CL. Colchicine decreased icROS in response to MSU crystals, while icROS production in response to PMA or S. aureus was not affected. Peak CL values are shown ( n = 7). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Article Snippet: After 5 min equilibration at 37 °C, cells were stimulated with MSU crystals (300 µg/mL, unless otherwise stated), PMA (50 nM), or serum-opsonized bacteria (Staphylococcus aureus opsonized with 10% human serum for 20 min at 37 °C) added at a multiplicity of infection (MOI) of 10, in the presence or absence of the following inhibitors or drugs; DPI (10 µM), GSK (20 µM) or cytochalasin B (5 µg/mL) at 37 °C for 5 min prior to use, or colchicine (1 µg/mL) at 37 °C for 20 min prior to use.

    Techniques: Amplification, Produced

    MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.

    Journal: International Journal of Molecular Sciences

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    doi: 10.3390/ijms21113750

    Figure Lengend Snippet: MSU crystals induce neutrophil extracellular trap (NET) formation. ( A ) Neutrophils stimulated with MSU crystals formed NETs as confirmed by DNA- (blue) and MPO- (green) labelled micrographs of neutrophils 3 h after stimulation with MSU crystals (300 µg/mL). Scale bar = 10 µm. ( B ) NET formation as measured with Sytox Green fluorescence over 3 h. To the left, a representative kinetic curve of neutrophils stimulated with buffer (black squares, broken line), MSU crystals (300 µg/mL, triangles, dotted line), or PMA (50 nM, black dots, solid line). MSU crystals (300 µg/mL) triggered a statistically significant DNA release ( p = 0.0005 compared to buffer treated cells after 180 min, n = 12). To the right, NET formation evaluated after 3 h with different concentrations of MSU crystals, mean fluorescence +/− SEM are shown ( n = 3). Statistical significance was calculated by the use of the Wilcoxon matched-pairs signed rank test.

    Article Snippet: After 5 min equilibration at 37 °C, cells were stimulated with MSU crystals (300 µg/mL, unless otherwise stated), PMA (50 nM), or serum-opsonized bacteria (Staphylococcus aureus opsonized with 10% human serum for 20 min at 37 °C) added at a multiplicity of infection (MOI) of 10, in the presence or absence of the following inhibitors or drugs; DPI (10 µM), GSK (20 µM) or cytochalasin B (5 µg/mL) at 37 °C for 5 min prior to use, or colchicine (1 µg/mL) at 37 °C for 20 min prior to use.

    Techniques: Fluorescence

    MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Journal: International Journal of Molecular Sciences

    Article Title: In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

    doi: 10.3390/ijms21113750

    Figure Lengend Snippet: MSU crystal induced NET formation is independent of ROS. ( A ) Neutrophils were pre-treated with the NADPH-oxidase inhibitors DPI (10 μM) or GSK (20 µM) and thereafter stimulated with MSU crystals (300 μg/mL) or PMA (50 nM) before NET formation was evaluated by Sytox green measurement after 3 h. As opposed to PMA stimulated neutrophils, MSU treated neutrophils formed NETs also in the presence of the inhibitors. Mean +/− SEM of three independent experiments are shown, a paired t -test was used for statistical comparisons. ( B ) Neutrophils from one patient with CGD (left) as well as from one individual with complete MPO-deficiency (MPO-, right) formed NETs in response to MSU (300 μg/mL), dotted lines, but not to PMA (50 nM, solid lines). * p ≤ 0.05 and ** p ≤ 0.01. ns = not significant.

    Article Snippet: After 5 min equilibration at 37 °C, cells were stimulated with MSU crystals (300 µg/mL, unless otherwise stated), PMA (50 nM), or serum-opsonized bacteria (Staphylococcus aureus opsonized with 10% human serum for 20 min at 37 °C) added at a multiplicity of infection (MOI) of 10, in the presence or absence of the following inhibitors or drugs; DPI (10 µM), GSK (20 µM) or cytochalasin B (5 µg/mL) at 37 °C for 5 min prior to use, or colchicine (1 µg/mL) at 37 °C for 20 min prior to use.

    Techniques:

    4-OI Reduces Inflammation in a Murine In Vivo Model of Peritonitis and Blocks NLRP3 Inflammasome Activation in Healthy Human and CAPS PBMCs (A–C) IL-1β concentration (A), IL-6 concentration (B), and neutrophil number (C) in the peritoneal lavage fluid of mice injected for 6 h with MSU crystals (30 mg/kg) ± 4-OI (50 mg/kg) (n = 3 for PBS groups, n = 8 for MSU groups). (D) LPS or Pam3CSK4 (14 h) and nigericin (2 h) induced IL-1β release (n = 5 for LPS + nigericin, n = 3 for Pam3CSK4 + nigericin) ± 4-OI or 4-O-2-MS (both 250 μM) from healthy human PBMCs. (E and F) Immunoblot analysis (E) and quantification by densitometry (F, n = 3) of pro- and mature IL-1β protein in lysates and supernatants of human PBMCs treated with LPS (14 h) and nigericin (2 h) ± 4-OI (250 μM). (G) LPS (1 h) induced IL-1β release (n = 3) ± 4-OI (250 μM) or MCC950 (500 nM) from PBMCs isolated from CAPS patients. ∗ p

    Journal: Cell Metabolism

    Article Title: The Immunomodulatory Metabolite Itaconate Modifies NLRP3 and Inhibits Inflammasome Activation

    doi: 10.1016/j.cmet.2020.07.016

    Figure Lengend Snippet: 4-OI Reduces Inflammation in a Murine In Vivo Model of Peritonitis and Blocks NLRP3 Inflammasome Activation in Healthy Human and CAPS PBMCs (A–C) IL-1β concentration (A), IL-6 concentration (B), and neutrophil number (C) in the peritoneal lavage fluid of mice injected for 6 h with MSU crystals (30 mg/kg) ± 4-OI (50 mg/kg) (n = 3 for PBS groups, n = 8 for MSU groups). (D) LPS or Pam3CSK4 (14 h) and nigericin (2 h) induced IL-1β release (n = 5 for LPS + nigericin, n = 3 for Pam3CSK4 + nigericin) ± 4-OI or 4-O-2-MS (both 250 μM) from healthy human PBMCs. (E and F) Immunoblot analysis (E) and quantification by densitometry (F, n = 3) of pro- and mature IL-1β protein in lysates and supernatants of human PBMCs treated with LPS (14 h) and nigericin (2 h) ± 4-OI (250 μM). (G) LPS (1 h) induced IL-1β release (n = 3) ± 4-OI (250 μM) or MCC950 (500 nM) from PBMCs isolated from CAPS patients. ∗ p

    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.

    Techniques: In Vivo, Activation Assay, Concentration Assay, Mouse Assay, Injection, Isolation

    MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P

    Journal: Arthritis Research & Therapy

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis

    doi: 10.1186/s13075-017-1418-6

    Figure Lengend Snippet: MicroRNA (miR, miRNA)-488 and miR-920 suppress monosodium urate (MSU)-induced expression of proinflammatory cytokines in THP-1 cells. The miRNA mimics or negative control (NC) mimics (50 nM) were transfected into THP-1 cells using Lipofectamine RNAiMAX reagent in accordance with the manufacturer’s instructions. After 24 h of transfection, cells were stimulated for 3 h with 0.5 μM 12-myristate 13-acetate. Then, cells were washed and stimulated with 250 μg/ml MSU crystals for 24 h to detect the production of proinflammatory cytokines. After the treatment, the cells were collected and analyzed by quantitative real-time polymerase chain reaction ( a , c ). The cell culture supernatants were also collected to detect the concentrations of interleukin (IL)-8 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay ( b , d ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. # P

    Article Snippet: MSU crystals can induce a variety of cytokines, including IL-1β, IL-6, IL-8, and TNF-α [ ].

    Techniques: Expressing, Negative Control, Transfection, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P

    Journal: Arthritis Research & Therapy

    Article Title: MicroRNA-488 and -920 regulate the production of proinflammatory cytokines in acute gouty arthritis

    doi: 10.1186/s13075-017-1418-6

    Figure Lengend Snippet: Monosodium urate (MSU) crystals promote the expression of proinflammatory cytokines in THP-1 cells. THP-1 cells were stimulated by the indicated concentration of MSU crystals. The messenger (mRNA) expression of interleukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α were detected by quantitative real-time polymerase chain reaction ( a – c ). Protein expression of IL-1β, IL-8, and TNF-α was detected by enzyme-linked immunosorbent assay ( d – f ). Values are expressed as mean ± SEM of three independent experiments, each of which was run in triplicate. * P

    Article Snippet: MSU crystals can induce a variety of cytokines, including IL-1β, IL-6, IL-8, and TNF-α [ ].

    Techniques: Expressing, Concentration Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    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