il 1β  (R&D Systems)

 
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    Name:
    Human IL 5 R alpha CD125 Antibody
    Description:
    The Human IL 5 R alpha CD125 Antibody from R D Systems is a goat polyclonal antibody to IL 5 R alpha CD125 This antibody reacts with human The Human IL 5 R alpha CD125 Antibody has been validated for the following applications Western Blot Flow Cytometry Neutralization CyTOF ready
    Catalog Number:
    AF-253-NA
    Price:
    369
    Category:
    Primary Antibodies
    Applications:
    Western Blot, Flow Cytometry, Neutralization, CyTOF-ready
    Purity:
    Immunogen affinity purified
    Conjugate:
    Unconjugated
    Immunogen:
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL-5 R alpha/CD125
    Size:
    100 ug
    Host:
    Goat
    Isotype:
    IgG
    Buy from Supplier


    Structured Review

    R&D Systems il 1β
    Human IL 5 R alpha CD125 Antibody
    The Human IL 5 R alpha CD125 Antibody from R D Systems is a goat polyclonal antibody to IL 5 R alpha CD125 This antibody reacts with human The Human IL 5 R alpha CD125 Antibody has been validated for the following applications Western Blot Flow Cytometry Neutralization CyTOF ready
    https://www.bioz.com/result/il 1β/product/R&D Systems
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 1β - by Bioz Stars, 2021-05
    86/100 stars

    Images

    1) Product Images from "Macrophage Activation in Acute Exacerbation of Idiopathic Pulmonary Fibrosis"

    Article Title: Macrophage Activation in Acute Exacerbation of Idiopathic Pulmonary Fibrosis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0116775

    Spontaneous macrophage derived chemokine production in IPF patients w/o acute exacerbation. Boxplots of the spontaneous production of A: CCL2, B: CCL17, C: CCL18, D: CCL22, E: IL-1ra, F: TNF-α, G: IL-1β, H: IL-8 and I: CXCL1 protein by BAL cells of patients with IPF. The dark grey boxplots represent patients who suffered from an acute exacerbation (AE, n = 12), the light grey represent patients who did’t suffer from an AE at timepoint of BAL (NoAE, n = 59) (* p
    Figure Legend Snippet: Spontaneous macrophage derived chemokine production in IPF patients w/o acute exacerbation. Boxplots of the spontaneous production of A: CCL2, B: CCL17, C: CCL18, D: CCL22, E: IL-1ra, F: TNF-α, G: IL-1β, H: IL-8 and I: CXCL1 protein by BAL cells of patients with IPF. The dark grey boxplots represent patients who suffered from an acute exacerbation (AE, n = 12), the light grey represent patients who did’t suffer from an AE at timepoint of BAL (NoAE, n = 59) (* p

    Techniques Used: Derivative Assay

    2) Product Images from "CpG-ODN and MPLA Prevent Mortality in a Murine Model of Post-Hemorrhage-Staphyloccocus aureus Pneumonia"

    Article Title: CpG-ODN and MPLA Prevent Mortality in a Murine Model of Post-Hemorrhage-Staphyloccocus aureus Pneumonia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0013228

    Whole Blood cells cultures. Hemorrhagic shock prior to methicillin-susceptible S. aureus (MSSA) pneumonia worsens peripherical blood reactivity after ex vivo LPS stimulation. Four groups of mice were studied: sham-treated (S group), hemorrhaged mice (H group); MSSA-induced pneumonia (P group), and hemorrhage before MSSA-induced pneumonia (HP group). Whole blood was exposed to LPS from E.coli O111 B4 for 24 hours and the following cytokines were assessed in the cell culture medium: ( A ) TNF-α, ( B ) IL-1β, and ( C ) MIP-2. Cytokine concentrations in the absence of LPS stimulation were always below the detection threshold (30 pg/ml). Data are representative of three independent experiments (each group, n = 6). Boxes represent median (interquartile range). € P
    Figure Legend Snippet: Whole Blood cells cultures. Hemorrhagic shock prior to methicillin-susceptible S. aureus (MSSA) pneumonia worsens peripherical blood reactivity after ex vivo LPS stimulation. Four groups of mice were studied: sham-treated (S group), hemorrhaged mice (H group); MSSA-induced pneumonia (P group), and hemorrhage before MSSA-induced pneumonia (HP group). Whole blood was exposed to LPS from E.coli O111 B4 for 24 hours and the following cytokines were assessed in the cell culture medium: ( A ) TNF-α, ( B ) IL-1β, and ( C ) MIP-2. Cytokine concentrations in the absence of LPS stimulation were always below the detection threshold (30 pg/ml). Data are representative of three independent experiments (each group, n = 6). Boxes represent median (interquartile range). € P

    Techniques Used: Ex Vivo, Mouse Assay, Cell Culture

    Hemorrhagic shock prior to methicillin-susceptible S. aureus (MSSA) pneumonia worsens lung damage. Lungs were harvested 24 hours after intratracheal instillation (S, P and HP groups) or hemorrhagic shock (H group). ( A, B, C, D ) Hematoxylin and eosin-stained sections of lungs (n = 3; magnification ×20) from mice that underwent the ( A ) sham procedure (S group), ( B ) Hemorrhage ( H ) , ( C ) MSSA-induced pneumonia (P group), or ( D ) hemorrhage before MSSA-induced pneumonia (HP group). Light microscopy revealed increased immune cell infiltration (purple stained) in group HP ( D ) compared with S, H and P groups. ( E ) Neutrophil accumulation assessed by myeloperoxidase activity in lung homogenates. ( F ) Vascular permeability was assessed in whole lung by measuring albumin-FITC passage through lung capillary. ( G ) Concentrations of TNF-α, IL-1β, and MIP-2 were assessed in lung homogenates. Data are representative of three independent experiments (each group, n = 6). Boxes represent median (interquartile range). P
    Figure Legend Snippet: Hemorrhagic shock prior to methicillin-susceptible S. aureus (MSSA) pneumonia worsens lung damage. Lungs were harvested 24 hours after intratracheal instillation (S, P and HP groups) or hemorrhagic shock (H group). ( A, B, C, D ) Hematoxylin and eosin-stained sections of lungs (n = 3; magnification ×20) from mice that underwent the ( A ) sham procedure (S group), ( B ) Hemorrhage ( H ) , ( C ) MSSA-induced pneumonia (P group), or ( D ) hemorrhage before MSSA-induced pneumonia (HP group). Light microscopy revealed increased immune cell infiltration (purple stained) in group HP ( D ) compared with S, H and P groups. ( E ) Neutrophil accumulation assessed by myeloperoxidase activity in lung homogenates. ( F ) Vascular permeability was assessed in whole lung by measuring albumin-FITC passage through lung capillary. ( G ) Concentrations of TNF-α, IL-1β, and MIP-2 were assessed in lung homogenates. Data are representative of three independent experiments (each group, n = 6). Boxes represent median (interquartile range). P

    Techniques Used: Staining, Mouse Assay, Light Microscopy, Activity Assay, Permeability

    3) Product Images from "HIV-1 Coinfection Profoundly Alters Intrahepatic Chemokine but Not Inflammatory Cytokine Profiles in HCV-Infected Subjects"

    Article Title: HIV-1 Coinfection Profoundly Alters Intrahepatic Chemokine but Not Inflammatory Cytokine Profiles in HCV-Infected Subjects

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086964

    Human hepatocytes produce CXCR3-associated chemokines in response to various stimulants. Huh7.5.1 cells in 24-well plates were treated or untreated with IFN-γ, IL-1β, TNF-α or their combinations at various concentrations for 24 h at 37°C. Cell-free supernatant and cells were subjected to measurement of expression of CXCL9, CXCL10, and CLCL11 using ELISA and FACS, respectively. A) IFN-γ induced Huh7.5.1 cells to produce CXCL9, CXCL10, and CXCL11 in a dose-dependent manner. B) FACS analysis of intracellular CXCL9, CXCL10, and CXCL11 levels in Huh7.5.1 cells in response to IFN-γ stimulation at 10 ng/mL. C) CXCL10, but not CXCL9 or CXCL11, was produced by Huh7.5.1 cells in response to IL-1β treatment in a dose-dependent manner. D) IL-1β, but not TNF-α, enhanced IFN-γ activity in induction of CXCR-associated chemokines by Huh7.5.1 cells. Error bars represent mean ± SD of values from triple wells in each dose. Note that different Y-axis values among CXCL9, CXCL10 and CXCL11 are used. NS, not significant; p ≥0.05; **, p
    Figure Legend Snippet: Human hepatocytes produce CXCR3-associated chemokines in response to various stimulants. Huh7.5.1 cells in 24-well plates were treated or untreated with IFN-γ, IL-1β, TNF-α or their combinations at various concentrations for 24 h at 37°C. Cell-free supernatant and cells were subjected to measurement of expression of CXCL9, CXCL10, and CLCL11 using ELISA and FACS, respectively. A) IFN-γ induced Huh7.5.1 cells to produce CXCL9, CXCL10, and CXCL11 in a dose-dependent manner. B) FACS analysis of intracellular CXCL9, CXCL10, and CXCL11 levels in Huh7.5.1 cells in response to IFN-γ stimulation at 10 ng/mL. C) CXCL10, but not CXCL9 or CXCL11, was produced by Huh7.5.1 cells in response to IL-1β treatment in a dose-dependent manner. D) IL-1β, but not TNF-α, enhanced IFN-γ activity in induction of CXCR-associated chemokines by Huh7.5.1 cells. Error bars represent mean ± SD of values from triple wells in each dose. Note that different Y-axis values among CXCL9, CXCL10 and CXCL11 are used. NS, not significant; p ≥0.05; **, p

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, FACS, Produced, Activity Assay

    Indirect effect of LPS on production of CXCR3-associated chemokines by human hepatocytes. A) LPS levels in plasma from subjected uninfected, or infected with HCV, HIV-1, or HCV/HIV-1 were determined using the QCL-1000 Chromogenic LAL assay. Lines in each plot represent mean ± SD of values from each group. B) Huh7.5.1 cells produced CXCL9, CXCL10 and CXCL11 in response to stimulation with 1∶10 diluted cell-free supernatant from LPS- or mock-treated pan T cells, monocytes, or PBMCs. IFN-γ at 1 ng/mL was used as a positive control of stimulation. Cell-free supernatant was subjected to ELISA assay to determine levels of CXCL9, CXCL10 and CXCL11. C) Levels of IFN-γ in the supernatant of monocytes or PBMCs treated with LPS or PBS were determined by the ELISA assay. D) LPS-S-mediated enhancement of CXCL9, CXCL10 and CXCL11 production by Huh7.5.1 cells was dependent on IFN-γ and IL-1β. LPS-S-mediated enhancement of CXCL9, CXCL10 and CXCL11 production by Huh7.5.1 cells was significantly blocked by IFN-γ nAb F12, and further down-regulated by IFN-γ nAb F12 (1 µg/mL) plus IL-1β nAb F2 (1 µg/mL). Horizontal bars represent mean ± SD of chemokine protein levels from triple wells of each dose. Note that different Y-axis values among CXCL9, CXCL10 and CXCL11 are used. LPS-S, supernatant from LPS-stimulated PBMCs. p
    Figure Legend Snippet: Indirect effect of LPS on production of CXCR3-associated chemokines by human hepatocytes. A) LPS levels in plasma from subjected uninfected, or infected with HCV, HIV-1, or HCV/HIV-1 were determined using the QCL-1000 Chromogenic LAL assay. Lines in each plot represent mean ± SD of values from each group. B) Huh7.5.1 cells produced CXCL9, CXCL10 and CXCL11 in response to stimulation with 1∶10 diluted cell-free supernatant from LPS- or mock-treated pan T cells, monocytes, or PBMCs. IFN-γ at 1 ng/mL was used as a positive control of stimulation. Cell-free supernatant was subjected to ELISA assay to determine levels of CXCL9, CXCL10 and CXCL11. C) Levels of IFN-γ in the supernatant of monocytes or PBMCs treated with LPS or PBS were determined by the ELISA assay. D) LPS-S-mediated enhancement of CXCL9, CXCL10 and CXCL11 production by Huh7.5.1 cells was dependent on IFN-γ and IL-1β. LPS-S-mediated enhancement of CXCL9, CXCL10 and CXCL11 production by Huh7.5.1 cells was significantly blocked by IFN-γ nAb F12, and further down-regulated by IFN-γ nAb F12 (1 µg/mL) plus IL-1β nAb F2 (1 µg/mL). Horizontal bars represent mean ± SD of chemokine protein levels from triple wells of each dose. Note that different Y-axis values among CXCL9, CXCL10 and CXCL11 are used. LPS-S, supernatant from LPS-stimulated PBMCs. p

    Techniques Used: Infection, Chromogenic LAL Assay, Produced, Positive Control, Enzyme-linked Immunosorbent Assay

    4) Product Images from "Pterostilbene reduces endothelial cell injury in vascular arterial walls by regulating the Nrf2-mediated AMPK/STAT3 pathway in an atherosclerosis rat model"

    Article Title: Pterostilbene reduces endothelial cell injury in vascular arterial walls by regulating the Nrf2-mediated AMPK/STAT3 pathway in an atherosclerosis rat model

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2019.8211

    Pts decreases the inflammatory response in an atherosclerosis rat model. Inflammatory cytokines (A) MCP-1, (B) IL-6, (C) IL-1β and (D) TNF-α in serum samples of Pts and PBS groups of rat models of atherosclerosis. **P
    Figure Legend Snippet: Pts decreases the inflammatory response in an atherosclerosis rat model. Inflammatory cytokines (A) MCP-1, (B) IL-6, (C) IL-1β and (D) TNF-α in serum samples of Pts and PBS groups of rat models of atherosclerosis. **P

    Techniques Used:

    5) Product Images from "A distal enhancer controls cytokine-dependent human cPLA2? gene expression [S]"

    Article Title: A distal enhancer controls cytokine-dependent human cPLA2? gene expression [S]

    Journal: Journal of Lipid Research

    doi: 10.1194/jlr.M037382

    Identification of transcription factors associated with the cPLA 2 α enhancer fragment. HFL cells were treated with IL-1β for the indicated times, followed by ChIP analysis as described in Materials and Methods. (A) ChIP analysis was performed
    Figure Legend Snippet: Identification of transcription factors associated with the cPLA 2 α enhancer fragment. HFL cells were treated with IL-1β for the indicated times, followed by ChIP analysis as described in Materials and Methods. (A) ChIP analysis was performed

    Techniques Used: Chromatin Immunoprecipitation

    IL-1β induction and chromatin structure of cPLA 2 α in HFL-1. (A, left) HFL-1 cells were treated with 2 ng/µl IL-1β for the indicated times, and total RNA was analyzed by real-time RT-PCR. Relative fold induction was calculated
    Figure Legend Snippet: IL-1β induction and chromatin structure of cPLA 2 α in HFL-1. (A, left) HFL-1 cells were treated with 2 ng/µl IL-1β for the indicated times, and total RNA was analyzed by real-time RT-PCR. Relative fold induction was calculated

    Techniques Used: Quantitative RT-PCR

    6) Product Images from "Aggravation of ovalbumin-induced murine asthma by co-exposure to desert-dust and organic chemicals: an animal model study"

    Article Title: Aggravation of ovalbumin-induced murine asthma by co-exposure to desert-dust and organic chemicals: an animal model study

    Journal: Environmental Health

    doi: 10.1186/1476-069X-13-83

    Expressions of IL-1β, IL-6, MCP-1 and MCP-3 in BALF. All values are expressed as mean ± SE (n =8). * p
    Figure Legend Snippet: Expressions of IL-1β, IL-6, MCP-1 and MCP-3 in BALF. All values are expressed as mean ± SE (n =8). * p

    Techniques Used:

    7) Product Images from "Reduced systemic and mycobacterial antigen – stimulated concentrations of IL-1β and IL-18 in Tuberculous Lymphadenitis"

    Article Title: Reduced systemic and mycobacterial antigen – stimulated concentrations of IL-1β and IL-18 in Tuberculous Lymphadenitis

    Journal: Cytokine

    doi: 10.1016/j.cyto.2016.10.013

    TBL is associated with reduced baseline, mycobacterial antigen and mitogen stimulated concentrations of IL-1β and IL-18 (A) The baseline or (B) mycobacterial antigen – stimulated or (C) mitogen stimulated concentrations of IFNγ, TNFα, IL-2, IL-1β, IL-18 and IL-10 were measured in TBL (n=31) and LTB (n=31) individuals. The results are shown as scatter plots with each circle representing a single individual and the bar representing the GM. The antigen or mitogen stimulated values are shown as net cytokine concentrations with the baseline subtracted. P values were calculated using the Mann-Whitney test with Holm’s correction for multiple comparisons.
    Figure Legend Snippet: TBL is associated with reduced baseline, mycobacterial antigen and mitogen stimulated concentrations of IL-1β and IL-18 (A) The baseline or (B) mycobacterial antigen – stimulated or (C) mitogen stimulated concentrations of IFNγ, TNFα, IL-2, IL-1β, IL-18 and IL-10 were measured in TBL (n=31) and LTB (n=31) individuals. The results are shown as scatter plots with each circle representing a single individual and the bar representing the GM. The antigen or mitogen stimulated values are shown as net cytokine concentrations with the baseline subtracted. P values were calculated using the Mann-Whitney test with Holm’s correction for multiple comparisons.

    Techniques Used: MANN-WHITNEY

    Treatment modifies the systemic cytokine profile in TBL (A) The systemic concentrations of Type 1 (IFNγ, TNFα, IL-2) and Type 17 (IL-17A, IL-17F, IL-22) cytokines were measured in TBL (n=31) individuals before (pre-T) and 6 months after (post-T) anti-TB treatment. (B) The systemic concentrations of pro-inflammatory (IL-1α, IL-1β, IL-12 and IL-18) and regulatory (IL-10, TGFβ) cytokines were measured in TBL (n=31) individuals before (pre-T) and 6 months after (post-T) anti-TB treatment. The results are shown as line diagrams with each line representing a single individual. P values were calculated using the Wilcoxon signed rank test with Holm’s correction for multiple comparisons.
    Figure Legend Snippet: Treatment modifies the systemic cytokine profile in TBL (A) The systemic concentrations of Type 1 (IFNγ, TNFα, IL-2) and Type 17 (IL-17A, IL-17F, IL-22) cytokines were measured in TBL (n=31) individuals before (pre-T) and 6 months after (post-T) anti-TB treatment. (B) The systemic concentrations of pro-inflammatory (IL-1α, IL-1β, IL-12 and IL-18) and regulatory (IL-10, TGFβ) cytokines were measured in TBL (n=31) individuals before (pre-T) and 6 months after (post-T) anti-TB treatment. The results are shown as line diagrams with each line representing a single individual. P values were calculated using the Wilcoxon signed rank test with Holm’s correction for multiple comparisons.

    Techniques Used:

    TBL is associated with reduced systemic concentrations of IL-1β and IL-18 (A) The systemic concentrations of Type 1 (IFNγ, TNFα, IL-2) and Type 17 (IL-17A, IL-17F, IL-22) cytokines were measured in TBL (n=31) and LTB (n=31) individuals. (B) The systemic concentrations of pro-inflammatory (IL-1α, IL-1β, IL-12 and IL-18) and regulatory (IL-10, TGFβ) cytokines were measured in TBL (n=31) and LTB (n=31) individuals. The results are shown as scatter plots with each circle representing a single individual and the bar representing the GM. P values were calculated using the Mann-Whitney test with Holm’s correction for multiple comparisons.
    Figure Legend Snippet: TBL is associated with reduced systemic concentrations of IL-1β and IL-18 (A) The systemic concentrations of Type 1 (IFNγ, TNFα, IL-2) and Type 17 (IL-17A, IL-17F, IL-22) cytokines were measured in TBL (n=31) and LTB (n=31) individuals. (B) The systemic concentrations of pro-inflammatory (IL-1α, IL-1β, IL-12 and IL-18) and regulatory (IL-10, TGFβ) cytokines were measured in TBL (n=31) and LTB (n=31) individuals. The results are shown as scatter plots with each circle representing a single individual and the bar representing the GM. P values were calculated using the Mann-Whitney test with Holm’s correction for multiple comparisons.

    Techniques Used: MANN-WHITNEY

    8) Product Images from "Helminth mediated modulation of the systemic and mycobacterial antigen – stimulated cytokine profiles in extra-pulmonary tuberculosis"

    Article Title: Helminth mediated modulation of the systemic and mycobacterial antigen – stimulated cytokine profiles in extra-pulmonary tuberculosis

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0007265

    TBL-Ss+ individuals exhibit elevated plasma levels of Type 17, Type 2 and diminished levels of pro-inflammatory cytokines. (A) The plasma levels of Type 1 (IFN-γ, TNF-α and IL-2), Type 17 (IL-17 and IL-22), (B) pro-inflammatory (IL-1α, IL-1β, IL-12, GM-CSF and IL-6), (C) Type 2 cytokines (IL-4, IL-5 and IL-13) and regulatory (IL-10) cytokines were examined by multiplex ELISA in TBL individuals with Strongyloides stercoralis (Ss) (Ss+, n = 44) or without Ss coinfection (Ss-, n = 44). The results are represented as scatterplots with each circle representing a single individual and the bar representing the geometric mean (GM). P values were calculated using the Mann-Whitney U test.
    Figure Legend Snippet: TBL-Ss+ individuals exhibit elevated plasma levels of Type 17, Type 2 and diminished levels of pro-inflammatory cytokines. (A) The plasma levels of Type 1 (IFN-γ, TNF-α and IL-2), Type 17 (IL-17 and IL-22), (B) pro-inflammatory (IL-1α, IL-1β, IL-12, GM-CSF and IL-6), (C) Type 2 cytokines (IL-4, IL-5 and IL-13) and regulatory (IL-10) cytokines were examined by multiplex ELISA in TBL individuals with Strongyloides stercoralis (Ss) (Ss+, n = 44) or without Ss coinfection (Ss-, n = 44). The results are represented as scatterplots with each circle representing a single individual and the bar representing the geometric mean (GM). P values were calculated using the Mann-Whitney U test.

    Techniques Used: Multiplex Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    TBL-Ss+ individuals exhibit diminished unstimulated and mycobacterial antigen stimulated levels of Type 1 and Type 17 cytokines. (A) The baseline/NIL (no antigen/negative control) or (B) mycobacterial antigen-stimulated or (C) mitogen stimulated levels of IFN-γ, TNF-α, IL-17, IL-22, IL-1α and IL-1β were examined by multiplex ELISA in Ss+ and Ss- individuals. The results are shown as scatterplots with each circle representing a single individual and the bar representing the GM. P values were calculated using the Mann-Whitney U test.
    Figure Legend Snippet: TBL-Ss+ individuals exhibit diminished unstimulated and mycobacterial antigen stimulated levels of Type 1 and Type 17 cytokines. (A) The baseline/NIL (no antigen/negative control) or (B) mycobacterial antigen-stimulated or (C) mitogen stimulated levels of IFN-γ, TNF-α, IL-17, IL-22, IL-1α and IL-1β were examined by multiplex ELISA in Ss+ and Ss- individuals. The results are shown as scatterplots with each circle representing a single individual and the bar representing the GM. P values were calculated using the Mann-Whitney U test.

    Techniques Used: Negative Control, Multiplex Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    9) Product Images from "Interleukin-1 beta induces the expression and production of stem cell factor by epithelial cells: crucial involvement of the PI-3K/mTOR pathway and HIF-1 transcription complex"

    Article Title: Interleukin-1 beta induces the expression and production of stem cell factor by epithelial cells: crucial involvement of the PI-3K/mTOR pathway and HIF-1 transcription complex

    Journal: Cellular and Molecular Immunology

    doi: 10.1038/cmi.2014.113

    The HIF-1 transcription complex is crucial for IL-1β-induced SCF expression in MCF-7 cells. ( a ) Normal and HIF-1α knockdown MCF-7 cells were exposed to 8 ng/ml IL-1β for 24 h followed by detection of intracellular HIF-1α/SCF levels as well as SCF release. ( b ) Normal and HIF-1α knockdown MCF-7 cells were exposed to 8 ng/ml IL-1β for 6 h followed by analysis of HIF-1α and SCF mRNA levels by qRT-PCR. Each sample was subjected to a contamination control through running qRT-PCR immediately after the decontamination procedure. No response was detected during 55 amplification cycles, indicating that contamination levels were close to zero (and are therefore not shown in the main figure). Western blot data show one representative experiment of 3–7 that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of 3–7 individual experiments; * P
    Figure Legend Snippet: The HIF-1 transcription complex is crucial for IL-1β-induced SCF expression in MCF-7 cells. ( a ) Normal and HIF-1α knockdown MCF-7 cells were exposed to 8 ng/ml IL-1β for 24 h followed by detection of intracellular HIF-1α/SCF levels as well as SCF release. ( b ) Normal and HIF-1α knockdown MCF-7 cells were exposed to 8 ng/ml IL-1β for 6 h followed by analysis of HIF-1α and SCF mRNA levels by qRT-PCR. Each sample was subjected to a contamination control through running qRT-PCR immediately after the decontamination procedure. No response was detected during 55 amplification cycles, indicating that contamination levels were close to zero (and are therefore not shown in the main figure). Western blot data show one representative experiment of 3–7 that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of 3–7 individual experiments; * P

    Techniques Used: Expressing, Quantitative RT-PCR, Amplification, Western Blot

    The PI-3K/mTOR pathway is crucially involved in IL-1β-induced HIF-1α accumulation in MCF-7 cells. Cells were pre-treated with inhibitors for 1 h (as indicated) and subsequently exposed to 8 ng/ml IL-1β for 4 h. The procedure was followed by Western blot analysis of HIF-1α accumulation ( a ) and detection of PI-3K and HIF-1α PHD activities as well as phospho-S2448 mTOR deposition ( b ). HIF-1α Western blot data show one representative experiment of three similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; results represent percentage values compared to the control. * P
    Figure Legend Snippet: The PI-3K/mTOR pathway is crucially involved in IL-1β-induced HIF-1α accumulation in MCF-7 cells. Cells were pre-treated with inhibitors for 1 h (as indicated) and subsequently exposed to 8 ng/ml IL-1β for 4 h. The procedure was followed by Western blot analysis of HIF-1α accumulation ( a ) and detection of PI-3K and HIF-1α PHD activities as well as phospho-S2448 mTOR deposition ( b ). HIF-1α Western blot data show one representative experiment of three similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; results represent percentage values compared to the control. * P

    Techniques Used: Western Blot

    IL-1β-induced SCF production by the MCF-7 cells is a time-dependent process. MCF-7 cells were exposed to 8 ng/ml IL-1β for different periods of time followed by analysis of intracellular HIF-1α and SCF accumulation (Western blot) and SCF release (ELISA). Western blot data show one representative experiment of three which demonstrated similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. for n =3; * P
    Figure Legend Snippet: IL-1β-induced SCF production by the MCF-7 cells is a time-dependent process. MCF-7 cells were exposed to 8 ng/ml IL-1β for different periods of time followed by analysis of intracellular HIF-1α and SCF accumulation (Western blot) and SCF release (ELISA). Western blot data show one representative experiment of three which demonstrated similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. for n =3; * P

    Techniques Used: Western Blot, Enzyme-linked Immunosorbent Assay

    IL-1β and SCF crosslinks using an in vitro coculture system and human plasma in vivo . ( a ) MCF-7 cells were cocultured with THP-1 cells at a ratio of 3∶1. Cocultures were exposed to 8 ng/ml IL-1β for 24 h in the absence or presence of 2 µg/ml human SCF neutralizing antibody which was added 30 min before IL-1β. ( b ) MTS cell viability and in-cell assay of SCF binding to THP-1 cells and VEGF release as outlined in Materials and Methods. ( c ) IL-1β and SCF levels were analysed in the plasma of ten healthy human donors by ELISA. Correlation analysis was performed using MS Excel software. Imaging data show one representative experiment of three that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; * P
    Figure Legend Snippet: IL-1β and SCF crosslinks using an in vitro coculture system and human plasma in vivo . ( a ) MCF-7 cells were cocultured with THP-1 cells at a ratio of 3∶1. Cocultures were exposed to 8 ng/ml IL-1β for 24 h in the absence or presence of 2 µg/ml human SCF neutralizing antibody which was added 30 min before IL-1β. ( b ) MTS cell viability and in-cell assay of SCF binding to THP-1 cells and VEGF release as outlined in Materials and Methods. ( c ) IL-1β and SCF levels were analysed in the plasma of ten healthy human donors by ELISA. Correlation analysis was performed using MS Excel software. Imaging data show one representative experiment of three that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; * P

    Techniques Used: In Vitro, In Vivo, Binding Assay, Enzyme-linked Immunosorbent Assay, Mass Spectrometry, Software, Imaging

    Crucial involvement of the PI-3K/mTOR pathway and HIF-1 transcription complex in IL-1β induced SCF production and secretion. eIF4E, eukaryotic translation initiation factor 4E; HIF, hypoxia-inducible factor; mTOR, mammalian target of rapamycin; PDK, phosphatidylinositol (P-3-PI)-dependent kinase; PI-3K, phosphatidylinositol-3 kinase; SCF, stem cell factor; S6K1, ribosomal protein S6 kinase beta-1; TSC, tuberous sclerosis complex.
    Figure Legend Snippet: Crucial involvement of the PI-3K/mTOR pathway and HIF-1 transcription complex in IL-1β induced SCF production and secretion. eIF4E, eukaryotic translation initiation factor 4E; HIF, hypoxia-inducible factor; mTOR, mammalian target of rapamycin; PDK, phosphatidylinositol (P-3-PI)-dependent kinase; PI-3K, phosphatidylinositol-3 kinase; SCF, stem cell factor; S6K1, ribosomal protein S6 kinase beta-1; TSC, tuberous sclerosis complex.

    Techniques Used:

    The level of IL-1β-induced HIF-1α accumulation in MCF-7 cells is comparable with those observed for classic HIF-1 activators. MCF-7 cells were exposed for 4 h to 8 ng/ml IL-1β, hypoxia (3% oxygen), cobalt chloride (100 µM) or MG132 (5 µM) followed by detection of HIF-1α accumulation and HIF-1 DBA as outlined in the ‘Materials and methods' section. Western blot data show one representative experiment of three that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of n =3; * P
    Figure Legend Snippet: The level of IL-1β-induced HIF-1α accumulation in MCF-7 cells is comparable with those observed for classic HIF-1 activators. MCF-7 cells were exposed for 4 h to 8 ng/ml IL-1β, hypoxia (3% oxygen), cobalt chloride (100 µM) or MG132 (5 µM) followed by detection of HIF-1α accumulation and HIF-1 DBA as outlined in the ‘Materials and methods' section. Western blot data show one representative experiment of three that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of n =3; * P

    Techniques Used: Western Blot

    IL-1β induces SCF production and HIF-1α accumulation in MCF-7 cells in a concentration-dependent manner. ( a ) MCF-7 cells were exposed to increasing concentrations of IL-1β for 24 h followed by detection of SCF release and HIF-1α accumulation. ( b ) THP-1 human myeloid leukaemia cells were treated in the same way and were used as a negative control cell line which does not produce SCF. Western blot data show one representative experiment of three that produced similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; * P
    Figure Legend Snippet: IL-1β induces SCF production and HIF-1α accumulation in MCF-7 cells in a concentration-dependent manner. ( a ) MCF-7 cells were exposed to increasing concentrations of IL-1β for 24 h followed by detection of SCF release and HIF-1α accumulation. ( b ) THP-1 human myeloid leukaemia cells were treated in the same way and were used as a negative control cell line which does not produce SCF. Western blot data show one representative experiment of three that produced similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; * P

    Techniques Used: Concentration Assay, Negative Control, Western Blot, Produced

    mTOR is crucially involved in IL-1β-induced SCF production. MCF-7 cells were pre-treated for 1 h with 10 µM rapamycin followed by 4 h exposure to 8 ng/ml IL-1β in the absence or presence of 50 µM CoCl 2 to maintain HIF-1 activity in the case of mTOR inhibition with rapamycin. HIF-1 DNA-binding activity and S2448 mTOR phosphorylation were monitored as described in the ‘Materials and methods' section. Western blot data show one representative experiment of three that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; * P
    Figure Legend Snippet: mTOR is crucially involved in IL-1β-induced SCF production. MCF-7 cells were pre-treated for 1 h with 10 µM rapamycin followed by 4 h exposure to 8 ng/ml IL-1β in the absence or presence of 50 µM CoCl 2 to maintain HIF-1 activity in the case of mTOR inhibition with rapamycin. HIF-1 DNA-binding activity and S2448 mTOR phosphorylation were monitored as described in the ‘Materials and methods' section. Western blot data show one representative experiment of three that gave similar results and were quantitatively analysed. Quantitative data are shown as mean values±s.d. of at least three individual experiments; * P

    Techniques Used: Activity Assay, Inhibition, Binding Assay, Western Blot

    10) Product Images from "A novel danshensu derivative ameliorates experimental colitis by modulating NADPH oxidase 4‐dependent NLRP3 inflammasome activation, et al. A novel danshensu derivative ameliorates experimental colitis by modulating NADPH oxidase 4‐dependent NLRP3 inflammasome activation"

    Article Title: A novel danshensu derivative ameliorates experimental colitis by modulating NADPH oxidase 4‐dependent NLRP3 inflammasome activation, et al. A novel danshensu derivative ameliorates experimental colitis by modulating NADPH oxidase 4‐dependent NLRP3 inflammasome activation

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.15890

    DSC inhibits the activation of NLRP3 inflammasome in vivo and in vitro. A, Colitis was induced as described in Materials and Methods and treated with or without DSC (50 mg/kg). Representative bands and quantitative analyses of NLRP3, caspase‐1 p20, cleaved IL‐1β and cleaved IL‐18 in colonic tissues. The β‐actin was used as loading control (n = 8 in each group). B, Colitis was induced as described in Materials and Methods and treated with or without lentiviral Nox4 shRNA. Representative bands and quantitative analyses of NLRP3, caspase‐1 p20, cleaved IL‐1β and cleaved IL‐18 in colonic tissues. The β‐actin was used as loading control (n = 8 in each group). C, NLRP3 inflammasome was induced in BMDM as described in Materials and Methods and treated with DSC (50 μmol/L). Representative bands of NLRP3, ASC and caspase‐1 in BMDM and caspase‐1 p20 in supernatant (SN) by Western blot and quantitative analyses of IL‐1β in supernatant by ELISA. The β‐actin was used as loading control. Data shown are means ± SEM of n = 8 in each group. *P
    Figure Legend Snippet: DSC inhibits the activation of NLRP3 inflammasome in vivo and in vitro. A, Colitis was induced as described in Materials and Methods and treated with or without DSC (50 mg/kg). Representative bands and quantitative analyses of NLRP3, caspase‐1 p20, cleaved IL‐1β and cleaved IL‐18 in colonic tissues. The β‐actin was used as loading control (n = 8 in each group). B, Colitis was induced as described in Materials and Methods and treated with or without lentiviral Nox4 shRNA. Representative bands and quantitative analyses of NLRP3, caspase‐1 p20, cleaved IL‐1β and cleaved IL‐18 in colonic tissues. The β‐actin was used as loading control (n = 8 in each group). C, NLRP3 inflammasome was induced in BMDM as described in Materials and Methods and treated with DSC (50 μmol/L). Representative bands of NLRP3, ASC and caspase‐1 in BMDM and caspase‐1 p20 in supernatant (SN) by Western blot and quantitative analyses of IL‐1β in supernatant by ELISA. The β‐actin was used as loading control. Data shown are means ± SEM of n = 8 in each group. *P

    Techniques Used: Activation Assay, In Vivo, In Vitro, shRNA, Western Blot, Enzyme-linked Immunosorbent Assay

    11) Product Images from "Proinflammatory cytokines increase glial fibrillary acidic protein expression in enteric glia"

    Article Title: Proinflammatory cytokines increase glial fibrillary acidic protein expression in enteric glia

    Journal: Gut

    doi: 10.1136/gut.2003.012625

    Immunofluorescence staining for glial fibrillary acidic protein (GFAP) and S-100 revealed that interleukin 1β (IL-1β 80 ng/ml) increased the number of GFAP+ enteric glia cells (B) in comparison with the untreated control (A). Scale bars 50 μm.
    Figure Legend Snippet: Immunofluorescence staining for glial fibrillary acidic protein (GFAP) and S-100 revealed that interleukin 1β (IL-1β 80 ng/ml) increased the number of GFAP+ enteric glia cells (B) in comparison with the untreated control (A). Scale bars 50 μm.

    Techniques Used: Immunofluorescence, Staining

    (A) Percentage of glial fibrillary acidic protein positive (GFAP+) enteric glia in vivo and in vitro, after incubation with lipopolysaccharide, the cytokines tumour necrosis factor α (TNF-α), interleukin (IL)-4, IL-1β, and anti-IL-1β/IL-1 receptor antagonist (ra), and after treatment with demecolcine. Data are mean (SEM) number of GFAP+ cells/S-100+ cells. *Significantly different from control (p
    Figure Legend Snippet: (A) Percentage of glial fibrillary acidic protein positive (GFAP+) enteric glia in vivo and in vitro, after incubation with lipopolysaccharide, the cytokines tumour necrosis factor α (TNF-α), interleukin (IL)-4, IL-1β, and anti-IL-1β/IL-1 receptor antagonist (ra), and after treatment with demecolcine. Data are mean (SEM) number of GFAP+ cells/S-100+ cells. *Significantly different from control (p

    Techniques Used: In Vivo, In Vitro, Incubation

    (A) Western blot analysis from cultured rat myenteric plexus cells (30 μg total protein loaded/lane). Lane 1: glial fibrillary acidic protein (GFAP) immunoreactivity of unstimulated cultures; lane 2: with 100 ng/ml interleukin (IL)-1β and 20 μg/ml anti-IL-1β; lane 3: with 5 ng/ml IL-1β; lane 4: with 10 ng/ml IL-1β; lane 5: with 20 ng/ml IL-1β; lane 6: with 80 ng/ml IL-1β; and lane 7: with 100 ng/ml IL-1β. (B) Western blot analysis from cultured rat myenteric plexus cells with demecolcine (30 μg total protein loaded/lane). Lane 1: GFAP amount of unstimulated cultures; lane 2: GFAP amount with demecolcine; lane 3: cultures stimulated with IL-1β (80 ng/ml); and lane 4: cultures after stimulation with IL-1β (80 ng/ml) and demecolcine.
    Figure Legend Snippet: (A) Western blot analysis from cultured rat myenteric plexus cells (30 μg total protein loaded/lane). Lane 1: glial fibrillary acidic protein (GFAP) immunoreactivity of unstimulated cultures; lane 2: with 100 ng/ml interleukin (IL)-1β and 20 μg/ml anti-IL-1β; lane 3: with 5 ng/ml IL-1β; lane 4: with 10 ng/ml IL-1β; lane 5: with 20 ng/ml IL-1β; lane 6: with 80 ng/ml IL-1β; and lane 7: with 100 ng/ml IL-1β. (B) Western blot analysis from cultured rat myenteric plexus cells with demecolcine (30 μg total protein loaded/lane). Lane 1: GFAP amount of unstimulated cultures; lane 2: GFAP amount with demecolcine; lane 3: cultures stimulated with IL-1β (80 ng/ml); and lane 4: cultures after stimulation with IL-1β (80 ng/ml) and demecolcine.

    Techniques Used: Western Blot, Cell Culture

    Effects of different concentrations of interleukin (IL)-1β on proliferation rates of primary glial fibrillary acidic protein positive (GFAP+) enteric glia by using bromodeoxyuridine (BrdU). Each bar represents the mean (SEM) of four independent experiments. There was no significant difference with respect to the proliferation rate of controls after incubation with IL-1β. Incorporation of BrdU was determined as 100% in controls.
    Figure Legend Snippet: Effects of different concentrations of interleukin (IL)-1β on proliferation rates of primary glial fibrillary acidic protein positive (GFAP+) enteric glia by using bromodeoxyuridine (BrdU). Each bar represents the mean (SEM) of four independent experiments. There was no significant difference with respect to the proliferation rate of controls after incubation with IL-1β. Incorporation of BrdU was determined as 100% in controls.

    Techniques Used: Incubation

    12) Product Images from "Carbohydrate response element binding protein (ChREBP) modulates the inflammatory response of mesangial cells in response to glucose"

    Article Title: Carbohydrate response element binding protein (ChREBP) modulates the inflammatory response of mesangial cells in response to glucose

    Journal: Bioscience Reports

    doi: 10.1042/BSR20180767

    High glucose induced pro-inflammatory cytokines and apoptosis in mesangial cells ( A ) SV40 MES 13 cells were treated with 5, 15, and 50 mM glucose for 48 h, the levels of inflammatory cytokines (TNF-α, IL-1β, and IL-6) were determined by ELISA. ( B ) SV40 MES 13 cells were treated with 0, 5, 15, and 50 mM glucose for 48 h, cells were harvested and cell apoptosis ratio was analyzed by flow cytometry.
    Figure Legend Snippet: High glucose induced pro-inflammatory cytokines and apoptosis in mesangial cells ( A ) SV40 MES 13 cells were treated with 5, 15, and 50 mM glucose for 48 h, the levels of inflammatory cytokines (TNF-α, IL-1β, and IL-6) were determined by ELISA. ( B ) SV40 MES 13 cells were treated with 0, 5, 15, and 50 mM glucose for 48 h, cells were harvested and cell apoptosis ratio was analyzed by flow cytometry.

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

    Correlation between ChREBP and pro-inflammatory cytokines Correlation analysis between ChREBP and TNFα ( A ), IL-1β ( B ), and IL-6 ( C ).
    Figure Legend Snippet: Correlation between ChREBP and pro-inflammatory cytokines Correlation analysis between ChREBP and TNFα ( A ), IL-1β ( B ), and IL-6 ( C ).

    Techniques Used:

    ChREBP mediated inflammatory reactions in mesangial cells induced by glucose administration SV40 MES 13 cells were transfected with control siRNA or siRNA against ChREBP for 48 h, the mRNA ( A ) and protein ( B ) levels of ChREBP were determined by real-time PCR and Western blot assay, respectively. SV40 MES 13 cells were transfected with control siRNA or siRNA against ChREBP for 24 h and treated with 5, 15, and 50 mM glucose for additional 48 h, the levels of inflammatory cytokines including TNF-α ( C ), IL-1β ( D ), and IL-6 ( E ) were determined by ELISA; ** P
    Figure Legend Snippet: ChREBP mediated inflammatory reactions in mesangial cells induced by glucose administration SV40 MES 13 cells were transfected with control siRNA or siRNA against ChREBP for 48 h, the mRNA ( A ) and protein ( B ) levels of ChREBP were determined by real-time PCR and Western blot assay, respectively. SV40 MES 13 cells were transfected with control siRNA or siRNA against ChREBP for 24 h and treated with 5, 15, and 50 mM glucose for additional 48 h, the levels of inflammatory cytokines including TNF-α ( C ), IL-1β ( D ), and IL-6 ( E ) were determined by ELISA; ** P

    Techniques Used: Transfection, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

    Expression of serum ChREBP and pro-inflammatory cytokines in T2DM patients ELISA was performed to detect the production of inflammatory cytokines including TNFα ( A ), IL-1β ( B ) and IL-6 ( C ), and ChREBP ( D );** P
    Figure Legend Snippet: Expression of serum ChREBP and pro-inflammatory cytokines in T2DM patients ELISA was performed to detect the production of inflammatory cytokines including TNFα ( A ), IL-1β ( B ) and IL-6 ( C ), and ChREBP ( D );** P

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay

    13) Product Images from "Mitochondrial ROS promote macrophage pyroptosis by inducing GSDMD oxidation"

    Article Title: Mitochondrial ROS promote macrophage pyroptosis by inducing GSDMD oxidation

    Journal: Journal of Molecular Cell Biology

    doi: 10.1093/jmcb/mjz020

    Redox status affects the cleavage of Gsdmd. ( A , D , and G ) Quantification of cell death via LDH release assay. ( B ) Immunoblot analysis of Gsdmd cleavage, caspase-1 activation, and IL-1β in cell lysates and supernatants of J774A.1 cells treated with gradient NAC (5, 10, 15, and 20 mM) and then stimulated with nigericin (10 μM). ( C ) The greyscale analysis of Gsdmd-N terminal in cell lysate (Input) and caspase-1 p20 subunit in both cell lysate (Input, black columns) and supernatant (SN, white columns) with loading control (β-actin), consisted with Figure 3 B . ( E ) Primed J774A.1 cells were given gradient NAC (10, 15 and 20 mM) preincubation. Cell lysates and supernatants were collected for immunoblot analysis of Gsdmd cleavage, caspase-1 activation, and IL-1β at 1 h after poly (dA:dT) (2 μg/ml) transfection. ( F ) The greyscale analysis of Gsdmd-N terminal in cell lysate (Input) and caspase-1 p20 subunit in both cell lysate (Input, black columns) and supernatant (SN, white columns) with loading control (α-tubulin), consisted with Figure 3 E . ( H ) LPS-primed J774A.1 cells were prestimulated with gradient 3-MA (1, 2.5, 5, and 10 mM) before the end of the priming process. Gsdmd cleavage, caspase-1 activation, and IL-1β were analyzed 1 h after nigericin stimulation via western blot. ( I ) The greyscale analysis of Gsdmd-N terminal in cell lysate (Input) and caspase-1 p20 subunit in both cell lysate (Input, black columns) and supernatant (SN, white columns) with loading control (β-actin), consisted with Figure 3 H . ( J ) Immunoblot analysis of the cleavage of human GSDMD and mouse Gsdmd by active CASP1 subunits after gradient H 2 O 2 (100 and 200 μM) incubation. ( K ) LDH assay of cell death in 293T cells expressing GSDMD/Gsdmd and active CASP1 subunits treated with gradient H 2 O 2 or left untreated. SN, supernatant; Input, whole-cell lysate; OE, overexpression; clea., cleavage. Data represent the average of three measurements. Mean ± SD of three experiments is shown. * P
    Figure Legend Snippet: Redox status affects the cleavage of Gsdmd. ( A , D , and G ) Quantification of cell death via LDH release assay. ( B ) Immunoblot analysis of Gsdmd cleavage, caspase-1 activation, and IL-1β in cell lysates and supernatants of J774A.1 cells treated with gradient NAC (5, 10, 15, and 20 mM) and then stimulated with nigericin (10 μM). ( C ) The greyscale analysis of Gsdmd-N terminal in cell lysate (Input) and caspase-1 p20 subunit in both cell lysate (Input, black columns) and supernatant (SN, white columns) with loading control (β-actin), consisted with Figure 3 B . ( E ) Primed J774A.1 cells were given gradient NAC (10, 15 and 20 mM) preincubation. Cell lysates and supernatants were collected for immunoblot analysis of Gsdmd cleavage, caspase-1 activation, and IL-1β at 1 h after poly (dA:dT) (2 μg/ml) transfection. ( F ) The greyscale analysis of Gsdmd-N terminal in cell lysate (Input) and caspase-1 p20 subunit in both cell lysate (Input, black columns) and supernatant (SN, white columns) with loading control (α-tubulin), consisted with Figure 3 E . ( H ) LPS-primed J774A.1 cells were prestimulated with gradient 3-MA (1, 2.5, 5, and 10 mM) before the end of the priming process. Gsdmd cleavage, caspase-1 activation, and IL-1β were analyzed 1 h after nigericin stimulation via western blot. ( I ) The greyscale analysis of Gsdmd-N terminal in cell lysate (Input) and caspase-1 p20 subunit in both cell lysate (Input, black columns) and supernatant (SN, white columns) with loading control (β-actin), consisted with Figure 3 H . ( J ) Immunoblot analysis of the cleavage of human GSDMD and mouse Gsdmd by active CASP1 subunits after gradient H 2 O 2 (100 and 200 μM) incubation. ( K ) LDH assay of cell death in 293T cells expressing GSDMD/Gsdmd and active CASP1 subunits treated with gradient H 2 O 2 or left untreated. SN, supernatant; Input, whole-cell lysate; OE, overexpression; clea., cleavage. Data represent the average of three measurements. Mean ± SD of three experiments is shown. * P

    Techniques Used: Lactate Dehydrogenase Assay, Activation Assay, Transfection, Western Blot, Incubation, Expressing, Over Expression

    The 4CS mutant of GSDMD blocks the protease activity of CASP1. ( A ) Co-immunoprecipitation analysis of the interaction of acCASP1 and GSDMD (WT, 4CS, D275A, and 4CS/D275A). ( B ) Exogenous CASP1 activity was analyzed following co-transfection with GSDMD (WT, 4CS, D275A, and 4CS/D275A). ( C ) Immunoblot analysis of the cleavage of human GSDMD and mouse Gsdmd (WT and 4CS) by active CASP1. ( D–G ) Overexpression of exogenous Gsdmd (WT and 4CS) in J774A.1 cells for 24 h that endogenous Gsdmd expression was downregulated by siRNA for 5 days. Then, Gsdmd cleavage and caspase-1 activation ( D and E ), cell death ( F ), and IL-1β release ( G ) were measured 1 h after Nlrp3 inflammasome activation. WT, wild-type; hGD, human GSDMD; mGd, mouse Gsdmd; simGd, mouse Gsdmd siRNA interference. Data represent the average of three measurements. Mean ± SD of three experiments is shown. * P
    Figure Legend Snippet: The 4CS mutant of GSDMD blocks the protease activity of CASP1. ( A ) Co-immunoprecipitation analysis of the interaction of acCASP1 and GSDMD (WT, 4CS, D275A, and 4CS/D275A). ( B ) Exogenous CASP1 activity was analyzed following co-transfection with GSDMD (WT, 4CS, D275A, and 4CS/D275A). ( C ) Immunoblot analysis of the cleavage of human GSDMD and mouse Gsdmd (WT and 4CS) by active CASP1. ( D–G ) Overexpression of exogenous Gsdmd (WT and 4CS) in J774A.1 cells for 24 h that endogenous Gsdmd expression was downregulated by siRNA for 5 days. Then, Gsdmd cleavage and caspase-1 activation ( D and E ), cell death ( F ), and IL-1β release ( G ) were measured 1 h after Nlrp3 inflammasome activation. WT, wild-type; hGD, human GSDMD; mGd, mouse Gsdmd; simGd, mouse Gsdmd siRNA interference. Data represent the average of three measurements. Mean ± SD of three experiments is shown. * P

    Techniques Used: Mutagenesis, Activity Assay, Immunoprecipitation, Cotransfection, Over Expression, Expressing, Activation Assay

    14) Product Images from "Citrullinated Histone H3 as a Therapeutic Target for Endotoxic Shock in Mice"

    Article Title: Citrullinated Histone H3 as a Therapeutic Target for Endotoxic Shock in Mice

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2019.02957

    The CitH3 mAb (4 Cit) improves survival and attenuates serum cytokines compared to the CitH3 mAb (3 Cit) in a mouse model of lethal endotoxic shock. C57BL/6J mice were randomized to injection: (1) IgG only (20 mg/kg), (2) LPS (20 mg/kg) + mouse IgG (20 mg/kg), LPS + CitH3 mAb (4 Cit) (~20 mg/kg), LPS + CitH3 mAb (3 Cit) (~20 mg/kg). (A) Survival was monitored for 10 days ( n = 9/group). Kaplan–Meier curves were used for survival rate analysis. The CitH3 mAb (4 Cit) significantly improved mouse survival compared to the LPS + mouse IgG group ( p = 0.0004) and to the LPS + CitH3 mAb (3 Cit) group ( p = 0.0039). There was no survival difference between the LPS + IgG group and the LPS + CitH3 mAb (3 Cit) group. (B) In another cohort ( n = 3/group), blood and organs were harvested at 12 h after LPS injection. Serum levels of IL-1β and TNF-α were measured using ELISA. Data are presented as mean ± SEM ( n = 3/group). * p
    Figure Legend Snippet: The CitH3 mAb (4 Cit) improves survival and attenuates serum cytokines compared to the CitH3 mAb (3 Cit) in a mouse model of lethal endotoxic shock. C57BL/6J mice were randomized to injection: (1) IgG only (20 mg/kg), (2) LPS (20 mg/kg) + mouse IgG (20 mg/kg), LPS + CitH3 mAb (4 Cit) (~20 mg/kg), LPS + CitH3 mAb (3 Cit) (~20 mg/kg). (A) Survival was monitored for 10 days ( n = 9/group). Kaplan–Meier curves were used for survival rate analysis. The CitH3 mAb (4 Cit) significantly improved mouse survival compared to the LPS + mouse IgG group ( p = 0.0004) and to the LPS + CitH3 mAb (3 Cit) group ( p = 0.0039). There was no survival difference between the LPS + IgG group and the LPS + CitH3 mAb (3 Cit) group. (B) In another cohort ( n = 3/group), blood and organs were harvested at 12 h after LPS injection. Serum levels of IL-1β and TNF-α were measured using ELISA. Data are presented as mean ± SEM ( n = 3/group). * p

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

    The CitH3 mAb (4 Cit) ameliorates acute lung injury in lethal endotoxic shock. Mice were randomly divided into four groups: (1) IgG only (20 mg/kg), (2) LPS (20 mg/kg) + IgG (20 mg/kg), (3) LPS + CitH3 mAb (4 Cit) (20 mg/kg), and (4) LPS + CitH3 mAb (3 Cit) (20 mg/kg) ( n = 3/group). Blood and organs were harvested at 12 h after LPS injection. (A) Representative hematoxylin and eosin staining of mouse lung sections is shown, and histological analysis of acute lung injury (ALI) was graded by a blinded pathologist and presented as ALI score (mean ± SEM, n = 3/group). White arrows indicate inflammatory changes. (B) IL-1β and TNF-α of lung homogenates were determined by ELISA. Cytokine levels were normalized by protein concentration and were significantly lower in CitH3 mAb (4 Cit)-treated mice (mean ± SEM, n = 3/group). * p
    Figure Legend Snippet: The CitH3 mAb (4 Cit) ameliorates acute lung injury in lethal endotoxic shock. Mice were randomly divided into four groups: (1) IgG only (20 mg/kg), (2) LPS (20 mg/kg) + IgG (20 mg/kg), (3) LPS + CitH3 mAb (4 Cit) (20 mg/kg), and (4) LPS + CitH3 mAb (3 Cit) (20 mg/kg) ( n = 3/group). Blood and organs were harvested at 12 h after LPS injection. (A) Representative hematoxylin and eosin staining of mouse lung sections is shown, and histological analysis of acute lung injury (ALI) was graded by a blinded pathologist and presented as ALI score (mean ± SEM, n = 3/group). White arrows indicate inflammatory changes. (B) IL-1β and TNF-α of lung homogenates were determined by ELISA. Cytokine levels were normalized by protein concentration and were significantly lower in CitH3 mAb (4 Cit)-treated mice (mean ± SEM, n = 3/group). * p

    Techniques Used: Mouse Assay, Injection, Staining, Enzyme-linked Immunosorbent Assay, Protein Concentration

    15) Product Images from "Nardostachys jatamansi inhibits severe acute pancreatitis via mitogen-activated protein kinases"

    Article Title: Nardostachys jatamansi inhibits severe acute pancreatitis via mitogen-activated protein kinases

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2012.612

    Effect of NJ on TNF-α, IL-1β and IL-6. To examine production of TNF-α, IL-1β and IL-6 (A) serum levels and (B) mRNA levels in the pancreas were measured by ELISA and real-time RT-PCR, respectively. Data are represented as the means ± SEM (n=6 in each group). * P
    Figure Legend Snippet: Effect of NJ on TNF-α, IL-1β and IL-6. To examine production of TNF-α, IL-1β and IL-6 (A) serum levels and (B) mRNA levels in the pancreas were measured by ELISA and real-time RT-PCR, respectively. Data are represented as the means ± SEM (n=6 in each group). * P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

    16) Product Images from "Alkaloids from Tetrastigma hemsleyanum and Their Anti-Inflammatory Effects on LPS-Induced RAW264.7 Cells"

    Article Title: Alkaloids from Tetrastigma hemsleyanum and Their Anti-Inflammatory Effects on LPS-Induced RAW264.7 Cells

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/molecules23061445

    Effects of APTH and compound 10 on LPS-stimulated PGE 2 , IL-1β and IL-6 production in RAW264.7 macrophages. Cells were preincubated for 1 h with or without the test compounds and then stimulated for 16 h with LPS (0.1 μg/mL). The PGE 2 ( A ), IL-1β ( B ), and IL-6 ( C ) levels were measured by ELISA. The results are expressed as the means ± standard error of the mean (SEM). Dexamethasone was used as the positive control. Values of * p
    Figure Legend Snippet: Effects of APTH and compound 10 on LPS-stimulated PGE 2 , IL-1β and IL-6 production in RAW264.7 macrophages. Cells were preincubated for 1 h with or without the test compounds and then stimulated for 16 h with LPS (0.1 μg/mL). The PGE 2 ( A ), IL-1β ( B ), and IL-6 ( C ) levels were measured by ELISA. The results are expressed as the means ± standard error of the mean (SEM). Dexamethasone was used as the positive control. Values of * p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Positive Control

    17) Product Images from "Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway"

    Article Title: Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway

    Journal: Journal of Clinical Medicine

    doi: 10.3390/jcm8122210

    Erastin suppresses the induction of inflammatory cytokines in LPS-induced bone marrow-derived macrophages (BMDMs). Transcription levels of ( A ) TNF-α and ( B ) IL-1β after activation of BMDMs with 1 μg/mL of LPS for 24 h with or without 5, 10, or 20 μM of erastin were measured using real-time qPCR. ( C ) Protein expression of inflammatory cytokines (TNF-α and IL-1β) in LPS-induced BMDMs with or without the indicated dose of erastin (1, 5, 10, or 20 μM) were analyzed by western blot. Cell lysates were harvested at 24 h after LPS treatment with or without indicated dose of erastin; β-actin was used as a loading control. ( D ) TNF-α, and ( E ) IL-1β secretion levels after treating BMDMs with 1 μg/mL of LPS for 24 h with or without erastin at various doses (5, 10, 20 μM) in culture medium were determined by ELISA assay. Time-dependent secretion of ( F ) TNF-α and ( G ) IL-1β from BMDMs following stimulation with 1 μg/mL of LPS for 8, 16, and 24 h with or without 20 μM of erastin was assayed using ELISA. Graphs represent mean of three independent experiments. Statistical analyses were performed using paired two-tailed Student’s t -test. * p
    Figure Legend Snippet: Erastin suppresses the induction of inflammatory cytokines in LPS-induced bone marrow-derived macrophages (BMDMs). Transcription levels of ( A ) TNF-α and ( B ) IL-1β after activation of BMDMs with 1 μg/mL of LPS for 24 h with or without 5, 10, or 20 μM of erastin were measured using real-time qPCR. ( C ) Protein expression of inflammatory cytokines (TNF-α and IL-1β) in LPS-induced BMDMs with or without the indicated dose of erastin (1, 5, 10, or 20 μM) were analyzed by western blot. Cell lysates were harvested at 24 h after LPS treatment with or without indicated dose of erastin; β-actin was used as a loading control. ( D ) TNF-α, and ( E ) IL-1β secretion levels after treating BMDMs with 1 μg/mL of LPS for 24 h with or without erastin at various doses (5, 10, 20 μM) in culture medium were determined by ELISA assay. Time-dependent secretion of ( F ) TNF-α and ( G ) IL-1β from BMDMs following stimulation with 1 μg/mL of LPS for 8, 16, and 24 h with or without 20 μM of erastin was assayed using ELISA. Graphs represent mean of three independent experiments. Statistical analyses were performed using paired two-tailed Student’s t -test. * p

    Techniques Used: Derivative Assay, Activation Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test

    Erastin inhibits NO metabolites (NO x ), TNF-α, and IL-1β production in a CLP-induced sepsis model. Time-dependent secretion of ( A ) NO metabolites, ( B ) TNF-α, and ( C ) IL-1β with or without erastin treatment after CLP were determined using ELISA assay in mouse serum samples. Mouse sera were collected at 0, 24, 48 h after CLP with or without erastin treatment. All groups, n = 5 Graph represents mean of five independent mouse serums level of indicated mediator and cytokines. Protein expression of iNOS in ( D ) lung, ( E ) liver, and ( F ) kidney tissues of sham and CLP-induced sepsis mice with or without erastin treatment were analyzed by western blot. All tissues were randomly collected from two mice of each group at 24 h after CLP with or without erastin administration; β-actin was used as a loading control; graphs represent relative band intensities. Statistical analyses were performed using paired two-tailed Student’s t -test. * p
    Figure Legend Snippet: Erastin inhibits NO metabolites (NO x ), TNF-α, and IL-1β production in a CLP-induced sepsis model. Time-dependent secretion of ( A ) NO metabolites, ( B ) TNF-α, and ( C ) IL-1β with or without erastin treatment after CLP were determined using ELISA assay in mouse serum samples. Mouse sera were collected at 0, 24, 48 h after CLP with or without erastin treatment. All groups, n = 5 Graph represents mean of five independent mouse serums level of indicated mediator and cytokines. Protein expression of iNOS in ( D ) lung, ( E ) liver, and ( F ) kidney tissues of sham and CLP-induced sepsis mice with or without erastin treatment were analyzed by western blot. All tissues were randomly collected from two mice of each group at 24 h after CLP with or without erastin administration; β-actin was used as a loading control; graphs represent relative band intensities. Statistical analyses were performed using paired two-tailed Student’s t -test. * p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, Mouse Assay, Western Blot, Two Tailed Test

    Erastin prevents septic death and downregulates NO metabolites, TNF-α, and IL-1β levels in a mouse model of LPS-induced endotoxemia. ( A ) Graph represents mouse survival rate after PBS and LPS injection (·i.p., 10 mg/kg) with or without erastin treatment (20 mg/kg); PBS (circle), LPS + DMSO (square), LPS + erastin (empty square); all groups, n = 10. ( B ) NO metabolites, ( C ) TNF-α, and ( D ) IL-1β secretion after the administration of LPS with or without erastin, as assessed using mouse serum samples. Mouse sera were collected at 24 h after PBS and LPS injection with or without erastin. All groups, n = 10. Graphs represent mean of 10 independent mouse serum levels of indicated mediator and cytokines. The expression levels of iNOS in ( E ) lung, ( F ) liver, and ( G ) kidney tissues were analyzed by western blotting after LPS injection with or without erastin treatment. All tissues were randomly collected from two mice of each group at 24 h after LPS injection with or without erastin administration; β-actin was used as a loading control; graphs are shown relative to band intensities. Statistical analyses were performed using paired two-tailed Student’s t -test. * p
    Figure Legend Snippet: Erastin prevents septic death and downregulates NO metabolites, TNF-α, and IL-1β levels in a mouse model of LPS-induced endotoxemia. ( A ) Graph represents mouse survival rate after PBS and LPS injection (·i.p., 10 mg/kg) with or without erastin treatment (20 mg/kg); PBS (circle), LPS + DMSO (square), LPS + erastin (empty square); all groups, n = 10. ( B ) NO metabolites, ( C ) TNF-α, and ( D ) IL-1β secretion after the administration of LPS with or without erastin, as assessed using mouse serum samples. Mouse sera were collected at 24 h after PBS and LPS injection with or without erastin. All groups, n = 10. Graphs represent mean of 10 independent mouse serum levels of indicated mediator and cytokines. The expression levels of iNOS in ( E ) lung, ( F ) liver, and ( G ) kidney tissues were analyzed by western blotting after LPS injection with or without erastin treatment. All tissues were randomly collected from two mice of each group at 24 h after LPS injection with or without erastin administration; β-actin was used as a loading control; graphs are shown relative to band intensities. Statistical analyses were performed using paired two-tailed Student’s t -test. * p

    Techniques Used: Injection, Expressing, Western Blot, Mouse Assay, Two Tailed Test

    18) Product Images from "Capsular Polysaccharide Is Involved in NLRP3 Inflammasome Activation by Klebsiella pneumoniae Serotype K1"

    Article Title: Capsular Polysaccharide Is Involved in NLRP3 Inflammasome Activation by Klebsiella pneumoniae Serotype K1

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00125-15

    K1-CPS induces IL-1β secretion through the NLRP3 inflammasome. (A to C) J774A.1 macrophages were incubated for 30 min with or without the NLRP3 inhibitor glybenclamide (Glyb), for 5.5 h with or without 1 μg/ml of K1-CPS, and for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (A), IL-1β in the culture medium (B), and TNF-α in the culture medium (C) were measured by Western blotting and ELISA, respectively. (D to F) J774A.1 macrophages stably transfected with a control shRNA plasmid (sh-SC), an NLRP3 shRNA plasmid (sh-NLRP3), and an ASC shRNA plasmid (sh-ASC) were incubated for 5.5 h with or without 1 μg/ml of K1-CPS or LPS and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (D), IL-1β in the culture medium (E), and TNF-α in the culture medium (F) were measured by Western blotting and ELISA, respectively. In panels A and D, the results are representative of three separate experiments. In panels B, C, E, and F, the data are expressed as means ± SD from three separate experiments. ***, P
    Figure Legend Snippet: K1-CPS induces IL-1β secretion through the NLRP3 inflammasome. (A to C) J774A.1 macrophages were incubated for 30 min with or without the NLRP3 inhibitor glybenclamide (Glyb), for 5.5 h with or without 1 μg/ml of K1-CPS, and for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (A), IL-1β in the culture medium (B), and TNF-α in the culture medium (C) were measured by Western blotting and ELISA, respectively. (D to F) J774A.1 macrophages stably transfected with a control shRNA plasmid (sh-SC), an NLRP3 shRNA plasmid (sh-NLRP3), and an ASC shRNA plasmid (sh-ASC) were incubated for 5.5 h with or without 1 μg/ml of K1-CPS or LPS and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (D), IL-1β in the culture medium (E), and TNF-α in the culture medium (F) were measured by Western blotting and ELISA, respectively. In panels A and D, the results are representative of three separate experiments. In panels B, C, E, and F, the data are expressed as means ± SD from three separate experiments. ***, P

    Techniques Used: Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Stable Transfection, Transfection, shRNA, Plasmid Preparation

    K1-CPS and K2-CPS induce IL-1β and IL-18 secretion through NLRP3 inflammasome in macrophages. (A and B) Human THP-1 macrophages were incubated for 30 min with or without NLRP3 inhibitor glybenclamide (Glyb), for 5.5 h with or without K1-CPS or K2-CPS, and then for 30 min with or without ATP or nigericin. The levels of IL-1β (A) and IL-18 (B) in the culture medium were measured by ELISA and Western blotting, respectively. (C) sh-SC and sh-NLRP3 J774A.1 cells were incubated for 5.5 h with or without K1-CPS or K2-CPS and then for 30 min with or without ATP or nigericin. The levels of IL-1β in the culture medium were measured by ELISA. In panels A and C, the data are expressed as the means ± SD from three separate experiments. In panel B, the results are representative of three separate experiments. ***, P
    Figure Legend Snippet: K1-CPS and K2-CPS induce IL-1β and IL-18 secretion through NLRP3 inflammasome in macrophages. (A and B) Human THP-1 macrophages were incubated for 30 min with or without NLRP3 inhibitor glybenclamide (Glyb), for 5.5 h with or without K1-CPS or K2-CPS, and then for 30 min with or without ATP or nigericin. The levels of IL-1β (A) and IL-18 (B) in the culture medium were measured by ELISA and Western blotting, respectively. (C) sh-SC and sh-NLRP3 J774A.1 cells were incubated for 5.5 h with or without K1-CPS or K2-CPS and then for 30 min with or without ATP or nigericin. The levels of IL-1β in the culture medium were measured by ELISA. In panels A and C, the data are expressed as the means ± SD from three separate experiments. In panel B, the results are representative of three separate experiments. ***, P

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

    Effect of mitochondria on K1-CPS-mediated NLRP3 inflammasome activation. (A to C) J774A.1 macrophages were incubated for 30 min with or without cyclosporine (Cyclos. A), for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (A), IL-1β in the culture medium (B), and TNF-α in the culture medium (C) were measured by Western blotting and ELISA, respectively. (D and E) J774A.1 macrophages were incubated for 30 min with or without Mito-TEMPO, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (D) and IL-1β in the culture medium (E) were measured by Western blotting and ELISA, respectively. In panels A and D, the results are representative of three separate experiments. In panels B, C, and E, the data are expressed as the means ± SD from three separate experiments. *, P
    Figure Legend Snippet: Effect of mitochondria on K1-CPS-mediated NLRP3 inflammasome activation. (A to C) J774A.1 macrophages were incubated for 30 min with or without cyclosporine (Cyclos. A), for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (A), IL-1β in the culture medium (B), and TNF-α in the culture medium (C) were measured by Western blotting and ELISA, respectively. (D and E) J774A.1 macrophages were incubated for 30 min with or without Mito-TEMPO, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (D) and IL-1β in the culture medium (E) were measured by Western blotting and ELISA, respectively. In panels A and D, the results are representative of three separate experiments. In panels B, C, and E, the data are expressed as the means ± SD from three separate experiments. *, P

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

    Effect of reactive oxygen species (ROS) on K1-CPS-induced NLRP3 inflammasome activation. (A) J774A.1 macrophages were incubated for 30 min with or without NAC or DPI, for 30 min with 2 μM 2′,7′-dichlorofluorescein diacetate, and then for 10 min with or without 1 μg/ml of K1-CPS. ROS generation in the cells was measured by 2′,7′-dichlorofluorescein diacetate. (B) J774A.1 macrophages were incubated for 30 min with or without NAC or DPI and then for 6 h with or without 1 μg/ml of K1-CPS. The levels of NLRP3 and pro-IL-1β in the cells were measured by Western blotting. (C and D) J774A.1 macrophages were incubated for 30 min with or without NAC, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (C) and IL-1β in the culture medium (D) were measured by Western blotting and ELISA, respectively. In panels A and D, the data are expressed as the means ± SD from three separate experiments. In panels B and C, the results are representative of three separate experiments. *, P
    Figure Legend Snippet: Effect of reactive oxygen species (ROS) on K1-CPS-induced NLRP3 inflammasome activation. (A) J774A.1 macrophages were incubated for 30 min with or without NAC or DPI, for 30 min with 2 μM 2′,7′-dichlorofluorescein diacetate, and then for 10 min with or without 1 μg/ml of K1-CPS. ROS generation in the cells was measured by 2′,7′-dichlorofluorescein diacetate. (B) J774A.1 macrophages were incubated for 30 min with or without NAC or DPI and then for 6 h with or without 1 μg/ml of K1-CPS. The levels of NLRP3 and pro-IL-1β in the cells were measured by Western blotting. (C and D) J774A.1 macrophages were incubated for 30 min with or without NAC, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (C) and IL-1β in the culture medium (D) were measured by Western blotting and ELISA, respectively. In panels A and D, the data are expressed as the means ± SD from three separate experiments. In panels B and C, the results are representative of three separate experiments. *, P

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

    K1-CPS induces IL-1β secretion through caspase-1. (A and B) J774A.1 macrophages were incubated for 5.5 h with or without K1-CPS or K1-CPS-TFA and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (A) and IL-1β in the culture medium (B) were measured by Western blotting and ELISA, respectively. (C and D) J774A.1 macrophages were incubated for 30 min with or without the caspase-1 inhibitor YVAD-CHO, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (C) and IL-1β in the culture medium (D) were measured by Western blotting and ELISA, respectively. In panels A and C, the results are representative of three separate experiments. In panels B and D, the data are expressed as the means ± SD from three separate experiments. ***, P
    Figure Legend Snippet: K1-CPS induces IL-1β secretion through caspase-1. (A and B) J774A.1 macrophages were incubated for 5.5 h with or without K1-CPS or K1-CPS-TFA and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (A) and IL-1β in the culture medium (B) were measured by Western blotting and ELISA, respectively. (C and D) J774A.1 macrophages were incubated for 30 min with or without the caspase-1 inhibitor YVAD-CHO, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (C) and IL-1β in the culture medium (D) were measured by Western blotting and ELISA, respectively. In panels A and C, the results are representative of three separate experiments. In panels B and D, the data are expressed as the means ± SD from three separate experiments. ***, P

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

    Effect of MAPK on K1-CPS-induced NLRP3 inflammasome activation. (A) J774A.1 macrophages were incubated for 30 min with or without MAPK inhibitors PD, SB, and SP and then for 15 min with or without K1-CPS. The phosphorylation levels of MAPK in the cells were measured by Western blotting. (B) J774A.1 macrophages were incubated for 30 min with or without MAPK inhibitors PD, SB, and SP and then for 6 h with or without K1-CPS. The levels of NLRP3 and pro-IL-1β in the cells were measured by Western blotting. (C and D) J774A.1 macrophages were incubated for 30 min with or without PD, SB, and SP, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (C) and IL-1β in the culture medium (D) were measured by Western blotting and ELISA, respectively. In panels A to C, the results are representative of three separate experiments. In panel D, the data are expressed as the means ± SD from three separate experiments. *, P
    Figure Legend Snippet: Effect of MAPK on K1-CPS-induced NLRP3 inflammasome activation. (A) J774A.1 macrophages were incubated for 30 min with or without MAPK inhibitors PD, SB, and SP and then for 15 min with or without K1-CPS. The phosphorylation levels of MAPK in the cells were measured by Western blotting. (B) J774A.1 macrophages were incubated for 30 min with or without MAPK inhibitors PD, SB, and SP and then for 6 h with or without K1-CPS. The levels of NLRP3 and pro-IL-1β in the cells were measured by Western blotting. (C and D) J774A.1 macrophages were incubated for 30 min with or without PD, SB, and SP, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (C) and IL-1β in the culture medium (D) were measured by Western blotting and ELISA, respectively. In panels A to C, the results are representative of three separate experiments. In panel D, the data are expressed as the means ± SD from three separate experiments. *, P

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

    K. pneumoniae infection induces IL-1β secretion partially through NLRP3 inflammasome. (A) J774A.1 macrophages infected with or without wild-type, magA mutant, wbbO mutant, or magA-wbbO mutant PLA K. pneumoniae for 24 h. The levels of IL-1β in the culture medium were measured by ELISA. (B to D) sh-SC and sh-NLRP3 (B), sh-SC and sh-NLRC4 (C), or sh-SC and sh-TLR4 (D) J774A.1 cells left uninfected or infected with wild-type PLA K. pneumoniae for 24 h. The levels of IL-1β in the culture medium were measured by ELISA. (E and F) sh-SC and sh-NLRP3 (E) or sh-SC and sh-NLRC4 (F) J774A.1 cells infected with or without wild-type PLA K. pneumoniae for 24 h. The cell viability was measured by alamarBlue assay. (G) sh-SC, sh-NLRP3, and sh-NLRC4 J774A.1 cells left uninfected or infected with wild-type PLA K. pneumoniae for 24 h. The cell viability was measured by cell cycle (sub-G 1 ) analysis using PI staining. (H) J774A.1 macrophages left uninfected or infected with wild-type, magA mutant, wbbO mutant, or magA-wbbO mutant PLA K. pneumoniae for 24 h. Cell viability was measured by alamarBlue assay. (I) J774A.1 macrophages were incubated for 24 h with or without K1-CPS. The cell viability was measured by alamarBlue assay. The data are expressed as the means ± SD from three separate experiments. NT, no treatment. *, P
    Figure Legend Snippet: K. pneumoniae infection induces IL-1β secretion partially through NLRP3 inflammasome. (A) J774A.1 macrophages infected with or without wild-type, magA mutant, wbbO mutant, or magA-wbbO mutant PLA K. pneumoniae for 24 h. The levels of IL-1β in the culture medium were measured by ELISA. (B to D) sh-SC and sh-NLRP3 (B), sh-SC and sh-NLRC4 (C), or sh-SC and sh-TLR4 (D) J774A.1 cells left uninfected or infected with wild-type PLA K. pneumoniae for 24 h. The levels of IL-1β in the culture medium were measured by ELISA. (E and F) sh-SC and sh-NLRP3 (E) or sh-SC and sh-NLRC4 (F) J774A.1 cells infected with or without wild-type PLA K. pneumoniae for 24 h. The cell viability was measured by alamarBlue assay. (G) sh-SC, sh-NLRP3, and sh-NLRC4 J774A.1 cells left uninfected or infected with wild-type PLA K. pneumoniae for 24 h. The cell viability was measured by cell cycle (sub-G 1 ) analysis using PI staining. (H) J774A.1 macrophages left uninfected or infected with wild-type, magA mutant, wbbO mutant, or magA-wbbO mutant PLA K. pneumoniae for 24 h. Cell viability was measured by alamarBlue assay. (I) J774A.1 macrophages were incubated for 24 h with or without K1-CPS. The cell viability was measured by alamarBlue assay. The data are expressed as the means ± SD from three separate experiments. NT, no treatment. *, P

    Techniques Used: Infection, Mutagenesis, Proximity Ligation Assay, Enzyme-linked Immunosorbent Assay, Alamar Blue Assay, Staining, Incubation

    19) Product Images from "Promyelocytic Leukemia Protein Interacts with the Apoptosis-associated Speck-like Protein to Limit Inflammasome Activation *"

    Article Title: Promyelocytic Leukemia Protein Interacts with the Apoptosis-associated Speck-like Protein to Limit Inflammasome Activation *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.539692

    PML-deficient BMDM display enhanced levels of IL-1β and IL-18 in response to NLRP3 and AIM2 inflammasome activation. Primary BMDM from 129Sv wild type ( WT ) and PML −/− mice were left unstimulated or stimulated with 100 ng/ml LPS
    Figure Legend Snippet: PML-deficient BMDM display enhanced levels of IL-1β and IL-18 in response to NLRP3 and AIM2 inflammasome activation. Primary BMDM from 129Sv wild type ( WT ) and PML −/− mice were left unstimulated or stimulated with 100 ng/ml LPS

    Techniques Used: Activation Assay, Mouse Assay

    PML deficiency enhances levels of IL-1β in response to HSV-1 and S. typhimurium and has no effect on caspase-1-dependent pyroptosis. A, for infection with HSV-1, BMDMs were left unstimulated and mock-infected or stimulated with LPS (100 ng/ml)
    Figure Legend Snippet: PML deficiency enhances levels of IL-1β in response to HSV-1 and S. typhimurium and has no effect on caspase-1-dependent pyroptosis. A, for infection with HSV-1, BMDMs were left unstimulated and mock-infected or stimulated with LPS (100 ng/ml)

    Techniques Used: Infection

    20) Product Images from "Squamous metaplasia amplifies pathologic epithelial-mesenchymal interactions in COPD patients"

    Article Title: Squamous metaplasia amplifies pathologic epithelial-mesenchymal interactions in COPD patients

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI32526

    IL-1β is coexpressed with integrins α v β 6 and α v β 8 in hyperproliferative suprabasal squamous metaplastic airway epithelium of COPD patients. Immunostaining of adjacent paraffin sections from an airway with a focus of SM (Sq. Met.: A , C , E , G , I , K , M , O , and Q ) compared with relatively normal airway mucosa ( B , D , F , H , J , L , N , P , and R ) from a COPD patient using antibodies to ( A and B ) IL-1β; ( C and D ) IL-1β preabsorbed with the immunogenic peptide (control); ( E and F ) the suprabasal markers anti-involucrin; ( G and H ) anti–keratin 6; ( I and J ) anti–keratin 14; ( K and L ) the TGF-β activating integrins β6; ( M and N ) and β8; ( O and P ) the cell proliferation marker Ki-67; ( Q and R ) the basal cell marker p63. Arrows point to the basement membrane. Shown is a representative experiment of 3 showing similar results. Scale bar: 100 μm.
    Figure Legend Snippet: IL-1β is coexpressed with integrins α v β 6 and α v β 8 in hyperproliferative suprabasal squamous metaplastic airway epithelium of COPD patients. Immunostaining of adjacent paraffin sections from an airway with a focus of SM (Sq. Met.: A , C , E , G , I , K , M , O , and Q ) compared with relatively normal airway mucosa ( B , D , F , H , J , L , N , P , and R ) from a COPD patient using antibodies to ( A and B ) IL-1β; ( C and D ) IL-1β preabsorbed with the immunogenic peptide (control); ( E and F ) the suprabasal markers anti-involucrin; ( G and H ) anti–keratin 6; ( I and J ) anti–keratin 14; ( K and L ) the TGF-β activating integrins β6; ( M and N ) and β8; ( O and P ) the cell proliferation marker Ki-67; ( Q and R ) the basal cell marker p63. Arrows point to the basement membrane. Shown is a representative experiment of 3 showing similar results. Scale bar: 100 μm.

    Techniques Used: Immunostaining, Marker

    IL-1α and IL-1β are induced during SM. ( A ) RT-PCR of total RNA from P0 to P3 human bronchial epithelial cells grown in 2D culture, using primers to IL-1α and IL-1β and β-actin, as a control. Shown is a representative experiment from paired samples from 3 different patients with the same results. ( B ) Antibody cytokine array showing increased IL-1β and -α in conditioned medium taken from P0 or P3 human bronchial epithelial cells. Upper and lower panels are duplicates. Shown is a representative experiment from 3 separate patients showing similar results. ( C ) IL-1β ELISA assay showing increased IL-1β in conditioned medium from P0 compared with P3 human bronchial epithelial cells. ** P
    Figure Legend Snippet: IL-1α and IL-1β are induced during SM. ( A ) RT-PCR of total RNA from P0 to P3 human bronchial epithelial cells grown in 2D culture, using primers to IL-1α and IL-1β and β-actin, as a control. Shown is a representative experiment from paired samples from 3 different patients with the same results. ( B ) Antibody cytokine array showing increased IL-1β and -α in conditioned medium taken from P0 or P3 human bronchial epithelial cells. Upper and lower panels are duplicates. Shown is a representative experiment from 3 separate patients showing similar results. ( C ) IL-1β ELISA assay showing increased IL-1β in conditioned medium from P0 compared with P3 human bronchial epithelial cells. ** P

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    Paracrine secretion of IL-1β by P3 human bronchial epithelial cells increases β8 expression and α v β 8 -mediated TGF-β activation of adjacent airway fibroblasts in a coculture model of the epithelial-mesenchymal trophic unit. Normal adult airway fibroblasts were cultured alone (Mono) or cocultured (Co) with P3 human bronchial epithelial cells grown on filter inserts. ( A ) Total fibroblast RNA was harvested and RT-PCR performed using primers specific for β8 or β-actin, as a control. Shown is a representative experiment of 3 with similar results. No cDNA indicates a no template control. ( B ) Fibroblasts ( n = 3) in mono- or coculture were stained with anti-β8 and analyzed by flow cytometry. Shown is mean fluorescence in arbitrary units. ( C ) Monocultured fibroblasts or fibroblasts after 48-hour coculture with P0 or P3 human bronchial epithelial cells ( n = 6) were cocultured with TGF-β reporter cells in the presence of neutralizing anti-β8, or no antibody (none). Shown is fold increase in TGF-β activation relative to untreated fibroblasts in monoculture. ( D ) Airway fibroblasts ( n = 3) were cocultured with P0 or P3 conditioned medium (CM) in the presence of no antagonist or different concentrations (5, 50, or 500 ng/ml) of IL-1RA, a naturally occurring soluble antagonist of IL-1α and IL-1β. The fibroblasts were stained with anti-β8 and analyzed using flow cytometry. Shown is the fold increase in β8 expression compared with matched airway fibroblasts grown without conditioned medium. ** P
    Figure Legend Snippet: Paracrine secretion of IL-1β by P3 human bronchial epithelial cells increases β8 expression and α v β 8 -mediated TGF-β activation of adjacent airway fibroblasts in a coculture model of the epithelial-mesenchymal trophic unit. Normal adult airway fibroblasts were cultured alone (Mono) or cocultured (Co) with P3 human bronchial epithelial cells grown on filter inserts. ( A ) Total fibroblast RNA was harvested and RT-PCR performed using primers specific for β8 or β-actin, as a control. Shown is a representative experiment of 3 with similar results. No cDNA indicates a no template control. ( B ) Fibroblasts ( n = 3) in mono- or coculture were stained with anti-β8 and analyzed by flow cytometry. Shown is mean fluorescence in arbitrary units. ( C ) Monocultured fibroblasts or fibroblasts after 48-hour coculture with P0 or P3 human bronchial epithelial cells ( n = 6) were cocultured with TGF-β reporter cells in the presence of neutralizing anti-β8, or no antibody (none). Shown is fold increase in TGF-β activation relative to untreated fibroblasts in monoculture. ( D ) Airway fibroblasts ( n = 3) were cocultured with P0 or P3 conditioned medium (CM) in the presence of no antagonist or different concentrations (5, 50, or 500 ng/ml) of IL-1RA, a naturally occurring soluble antagonist of IL-1α and IL-1β. The fibroblasts were stained with anti-β8 and analyzed using flow cytometry. Shown is the fold increase in β8 expression compared with matched airway fibroblasts grown without conditioned medium. ** P

    Techniques Used: Expressing, Activation Assay, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Staining, Flow Cytometry, Cytometry, Fluorescence

    Global analysis of gene expression during SM reveals induction of the proinflammatory mediators IL-1α and IL-1β.
    Figure Legend Snippet: Global analysis of gene expression during SM reveals induction of the proinflammatory mediators IL-1α and IL-1β.

    Techniques Used: Expressing

    Hypothetical model of SM in the pathogenesis of airway wall thickening in COPD. ). (c) Squamous metaplastic epithelial cells secrete increased IL-1β, which acts a paracrine factor with adjacent airway fibroblasts. (d) Airway fibroblasts respond to IL-1β by increasing β8 expression and α v β 8 ) and contributes to the increased expression of β6 by adjacent airway epithelial cells, forming a self-amplifying loop of TGF-β activation.
    Figure Legend Snippet: Hypothetical model of SM in the pathogenesis of airway wall thickening in COPD. ). (c) Squamous metaplastic epithelial cells secrete increased IL-1β, which acts a paracrine factor with adjacent airway fibroblasts. (d) Airway fibroblasts respond to IL-1β by increasing β8 expression and α v β 8 ) and contributes to the increased expression of β6 by adjacent airway epithelial cells, forming a self-amplifying loop of TGF-β activation.

    Techniques Used: Expressing, Activation Assay

    21) Product Images from "Blocking Interleukin-1? Induces a Healing-Associated Wound Macrophage Phenotype and Improves Healing in Type 2 Diabetes"

    Article Title: Blocking Interleukin-1? Induces a Healing-Associated Wound Macrophage Phenotype and Improves Healing in Type 2 Diabetes

    Journal: Diabetes

    doi: 10.2337/db12-1450

    IL-1β–neutralizing antibody improves healing of wounds in diabetic mice. Images showing cryosections stained with trichrome for ( A ) control IgG-treated and ( B ) IL-1β–neutralizing antibody–treated wounds on day 10 postinjury. Note the increased re-epithelialization and granulation tissue in the IL-1β antibody (IL1ab)–treated wounds. Scale bar = 0.5 mm. Arrows indicate ends of migrating epithelial tongues. gt, granulation tissue; mm, deep muscle. The muscle layer observed underneath the wound in some sections is likely a part of the deep tissue collected that helps to maintain the integrity of the fragile wounds, particularly in untreated and IgG-treated diabetic mice. For wounds in db/+, untreated db/db, IgG-treated db/db, and IL1ab-treated db/db mice on day 10 postinjury, quantification of ( C , D ) re-epithelialization and granulation tissue thickness measured in hematoxylin and eosin–stained cryosections, ( E ) trichrome staining measured as percent area stained blue for collagen, and ( F ) CD31 staining measured as percent area stained for this endothelial cell marker. In addition, wounds from each group of mice were homogenized and levels of ( G–J ) IL-1β, TNF-α, IGF-1, and TGF-β measured using ELISA. For all graphs, bars = mean ± SD, n = 6–8. *Mean value significantly different from that for wounds in db/+ mice. **Mean value significantly different from that for control IgG-treated wounds.
    Figure Legend Snippet: IL-1β–neutralizing antibody improves healing of wounds in diabetic mice. Images showing cryosections stained with trichrome for ( A ) control IgG-treated and ( B ) IL-1β–neutralizing antibody–treated wounds on day 10 postinjury. Note the increased re-epithelialization and granulation tissue in the IL-1β antibody (IL1ab)–treated wounds. Scale bar = 0.5 mm. Arrows indicate ends of migrating epithelial tongues. gt, granulation tissue; mm, deep muscle. The muscle layer observed underneath the wound in some sections is likely a part of the deep tissue collected that helps to maintain the integrity of the fragile wounds, particularly in untreated and IgG-treated diabetic mice. For wounds in db/+, untreated db/db, IgG-treated db/db, and IL1ab-treated db/db mice on day 10 postinjury, quantification of ( C , D ) re-epithelialization and granulation tissue thickness measured in hematoxylin and eosin–stained cryosections, ( E ) trichrome staining measured as percent area stained blue for collagen, and ( F ) CD31 staining measured as percent area stained for this endothelial cell marker. In addition, wounds from each group of mice were homogenized and levels of ( G–J ) IL-1β, TNF-α, IGF-1, and TGF-β measured using ELISA. For all graphs, bars = mean ± SD, n = 6–8. *Mean value significantly different from that for wounds in db/+ mice. **Mean value significantly different from that for control IgG-treated wounds.

    Techniques Used: Mouse Assay, Staining, Marker, Enzyme-linked Immunosorbent Assay

    IL-1β–neutralizing antibody downregulates proinflammatory macrophage phenotype and upregulates healing-associated phenotype in wounds of diabetic mice. Wounds in db/db mice were treated with control IgG or IL-1β–blocking antibody (IL1ab) on day 3 postinjury and macrophages were isolated on day 5 (d5) and day 10 (d10) postinjury. Expression of proinflammatory markers ( A–D ) IL-1β, MMP-9, TNF-α, and IL-6 and healing-associated/anti-inflammatory markers ( E–H ) CD206, IGF-1, TGF-β, and IL-10 were measured by real-time PCR. For all graphs, bars = mean ± SD, n = 6. *Mean value significantly different from that for IgG-treated mice at same time point, P
    Figure Legend Snippet: IL-1β–neutralizing antibody downregulates proinflammatory macrophage phenotype and upregulates healing-associated phenotype in wounds of diabetic mice. Wounds in db/db mice were treated with control IgG or IL-1β–blocking antibody (IL1ab) on day 3 postinjury and macrophages were isolated on day 5 (d5) and day 10 (d10) postinjury. Expression of proinflammatory markers ( A–D ) IL-1β, MMP-9, TNF-α, and IL-6 and healing-associated/anti-inflammatory markers ( E–H ) CD206, IGF-1, TGF-β, and IL-10 were measured by real-time PCR. For all graphs, bars = mean ± SD, n = 6. *Mean value significantly different from that for IgG-treated mice at same time point, P

    Techniques Used: Mouse Assay, Blocking Assay, Isolation, Expressing, Real-time Polymerase Chain Reaction

    Macrophages isolated from wounds in diabetic mice exhibit a persistent proinflammatory phenotype that may be induced by the wound environment. Macrophages were isolated from wounds of nondiabetic (db/+) and diabetic (db/db) mice on days 5 and 10 postinjury, cultured overnight, and release of ( A–D) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. Also, bone marrow–derived macrophages from wild-type mice were cultured with conditioned medium (WCM) of day 5 (5d) or day 10 (10d) wounds from nondiabetic db/+ or diabetic db/db mice; expression of proinflammatory markers ( E–H ) IL-1β, MMP-9, TNF-α, and IL-6 and healing-associated/anti-inflammatory markers ( I–L ) CD206, IGF-1, TGF-β, and IL-10 were assessed by real-time PCR. In addition, release of ( M–P ) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. For all graphs, bars = mean ± SD. For in vivo experiments, n = 6–7 mice for each strain and time point. For in vitro experiments, a separate set of bone marrow–derived macrophages (each harvested from a different mouse) was used for each of two experiments and wound-conditioned medium was generated from three mice per experiment, with n = 6 for each strain and time point. *Mean value significantly different from that for same strain on day 5 postinjury. **Mean value for db/db significantly different from that for db/+ at same time point, P
    Figure Legend Snippet: Macrophages isolated from wounds in diabetic mice exhibit a persistent proinflammatory phenotype that may be induced by the wound environment. Macrophages were isolated from wounds of nondiabetic (db/+) and diabetic (db/db) mice on days 5 and 10 postinjury, cultured overnight, and release of ( A–D) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. Also, bone marrow–derived macrophages from wild-type mice were cultured with conditioned medium (WCM) of day 5 (5d) or day 10 (10d) wounds from nondiabetic db/+ or diabetic db/db mice; expression of proinflammatory markers ( E–H ) IL-1β, MMP-9, TNF-α, and IL-6 and healing-associated/anti-inflammatory markers ( I–L ) CD206, IGF-1, TGF-β, and IL-10 were assessed by real-time PCR. In addition, release of ( M–P ) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. For all graphs, bars = mean ± SD. For in vivo experiments, n = 6–7 mice for each strain and time point. For in vitro experiments, a separate set of bone marrow–derived macrophages (each harvested from a different mouse) was used for each of two experiments and wound-conditioned medium was generated from three mice per experiment, with n = 6 for each strain and time point. *Mean value significantly different from that for same strain on day 5 postinjury. **Mean value for db/db significantly different from that for db/+ at same time point, P

    Techniques Used: Isolation, Mouse Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, In Vivo, In Vitro, Generated

    Macrophages isolated from chronic wounds in diabetic patients exhibit a proinflammatory phenotype that may be induced by the wound environment. Macrophages were isolated from chronic wound biopsy specimens and expression of proinflammatory markers IL-1β, MMP-9, TNF-α, and IL-6 ( A–D ) and healing-associated/anti-inflammatory markers CD206, IGF-1, TGF-β, and IL-10 ( E–H) were assessed by real-time PCR. For comparison, blood-derived macrophages from healthy volunteers were left nonstimulated (Non), stimulated with TNF-α and IFN-γ (classically activated [CA]), or stimulated with chronic wound–conditioned medium (WCM). For the in vitro experiments, release of IL-1β, TNF-α, IGF-1, and TGF-β ( I–L ) into cell culture medium was measured by ELISA. In addition, chronic wound biopsy cryosections were immunostained for IL-1β and the macrophage marker CD68, with images taken at 20× ( M–O ; scale bar = 100 μm) and at 40× ( P–R ; scale bar = 50 μm); location of 40× images shown in box on 20× images. Nuclei stained with DAPI. For all graphs, bars = mean ± SD and n = 5 for both in vivo and in vitro experiments. *Mean value significantly different from that for nonstimulated controls, P
    Figure Legend Snippet: Macrophages isolated from chronic wounds in diabetic patients exhibit a proinflammatory phenotype that may be induced by the wound environment. Macrophages were isolated from chronic wound biopsy specimens and expression of proinflammatory markers IL-1β, MMP-9, TNF-α, and IL-6 ( A–D ) and healing-associated/anti-inflammatory markers CD206, IGF-1, TGF-β, and IL-10 ( E–H) were assessed by real-time PCR. For comparison, blood-derived macrophages from healthy volunteers were left nonstimulated (Non), stimulated with TNF-α and IFN-γ (classically activated [CA]), or stimulated with chronic wound–conditioned medium (WCM). For the in vitro experiments, release of IL-1β, TNF-α, IGF-1, and TGF-β ( I–L ) into cell culture medium was measured by ELISA. In addition, chronic wound biopsy cryosections were immunostained for IL-1β and the macrophage marker CD68, with images taken at 20× ( M–O ; scale bar = 100 μm) and at 40× ( P–R ; scale bar = 50 μm); location of 40× images shown in box on 20× images. Nuclei stained with DAPI. For all graphs, bars = mean ± SD and n = 5 for both in vivo and in vitro experiments. *Mean value significantly different from that for nonstimulated controls, P

    Techniques Used: Isolation, Expressing, Real-time Polymerase Chain Reaction, Derivative Assay, In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay, Marker, Staining, In Vivo

    IL-1β–neutralizing antibody downregulates diabetic wound–conditioned medium–induced proinflammatory phenotype and upregulates healing-associated phenotype in cultured macrophages. Bone marrow–derived macrophages from wild-type mice left nonstimulated (Non) or stimulated with day 10 db/db wound-conditioned medium (WCM) along with control IgG or stimulated with IL-1β–neutralizing antibody (ILab); expression of proinflammatory markers ( A–D ) IL-1β, MMP-9, TNF-α, and IL-6 and healing-associated/anti-inflammatory markers ( E–H ) CD206, IGF-1, TGF-β, and IL-10 were measured by real-time PCR. In addition, release of ( I–L ) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. For all graphs, bars = mean ± SD. For these experiments, a separate set of bone marrow–derived macrophages (each harvested from a different mouse) was used for each of two experiments and wound-conditioned medium was generated from three mice per experiment, with n = 6 for each condition. *Mean value significantly different from that for nonstimulated controls. **Mean value significantly different from that for conditioned medium plus IgG-treated samples, P
    Figure Legend Snippet: IL-1β–neutralizing antibody downregulates diabetic wound–conditioned medium–induced proinflammatory phenotype and upregulates healing-associated phenotype in cultured macrophages. Bone marrow–derived macrophages from wild-type mice left nonstimulated (Non) or stimulated with day 10 db/db wound-conditioned medium (WCM) along with control IgG or stimulated with IL-1β–neutralizing antibody (ILab); expression of proinflammatory markers ( A–D ) IL-1β, MMP-9, TNF-α, and IL-6 and healing-associated/anti-inflammatory markers ( E–H ) CD206, IGF-1, TGF-β, and IL-10 were measured by real-time PCR. In addition, release of ( I–L ) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. For all graphs, bars = mean ± SD. For these experiments, a separate set of bone marrow–derived macrophages (each harvested from a different mouse) was used for each of two experiments and wound-conditioned medium was generated from three mice per experiment, with n = 6 for each condition. *Mean value significantly different from that for nonstimulated controls. **Mean value significantly different from that for conditioned medium plus IgG-treated samples, P

    Techniques Used: Cell Culture, Derivative Assay, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Generated

    Cultured macrophages from IL-1 receptor 1 knockout (IL1R1ko) mice are less sensitive to diabetic wound–conditioned medium than macrophages from wild-type mice. Bone marrow–derived macrophages from wild-type (WT) and IL1R1ko mice stimulated with day 10 db/db wound–conditioned medium, expression of proinflammatory markers ( A–D ) IL-1β, MMP-9, TNF-α, and IL-6, and healing-associated/anti-inflammatory markers ( E–H ) CD206, IGF-1, TGF-β, and IL-10 were measured by real-time PCR. In addition, release of ( I–L ) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. For all graphs, bars = mean ± SD. For these experiments, a separate set of bone marrow–derived macrophages (each harvested from a different mouse) was used for each of two experiments and wound-conditioned medium was generated from three mice per experiment, with n = 6 for each condition. *Mean value significantly different from that for nonstimulated controls of same strain. **Mean value significantly different from that for wound-conditioned medium–treated wild-type macrophages, P
    Figure Legend Snippet: Cultured macrophages from IL-1 receptor 1 knockout (IL1R1ko) mice are less sensitive to diabetic wound–conditioned medium than macrophages from wild-type mice. Bone marrow–derived macrophages from wild-type (WT) and IL1R1ko mice stimulated with day 10 db/db wound–conditioned medium, expression of proinflammatory markers ( A–D ) IL-1β, MMP-9, TNF-α, and IL-6, and healing-associated/anti-inflammatory markers ( E–H ) CD206, IGF-1, TGF-β, and IL-10 were measured by real-time PCR. In addition, release of ( I–L ) IL-1β, TNF-α, IGF-1, and TGF-β was measured using ELISA. For all graphs, bars = mean ± SD. For these experiments, a separate set of bone marrow–derived macrophages (each harvested from a different mouse) was used for each of two experiments and wound-conditioned medium was generated from three mice per experiment, with n = 6 for each condition. *Mean value significantly different from that for nonstimulated controls of same strain. **Mean value significantly different from that for wound-conditioned medium–treated wild-type macrophages, P

    Techniques Used: Cell Culture, Knock-Out, Mouse Assay, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Generated

    22) Product Images from "IPAF inflammasome is involved in IL-1β production from astrocytes, induced by palmitate; implications for Alzheimer’s Disease"

    Article Title: IPAF inflammasome is involved in IL-1β production from astrocytes, induced by palmitate; implications for Alzheimer’s Disease

    Journal: Neurobiology of aging

    doi: 10.1016/j.neurobiolaging.2013.08.016

    Pro-IL-1β and IL-1β levels produced by astrocytes with PA treatment
    Figure Legend Snippet: Pro-IL-1β and IL-1β levels produced by astrocytes with PA treatment

    Techniques Used: Produced

    The upregulated caspase-1 increased IL-1β level
    Figure Legend Snippet: The upregulated caspase-1 increased IL-1β level

    Techniques Used:

    IPAF is involved in IL-1β maturation
    Figure Legend Snippet: IPAF is involved in IL-1β maturation

    Techniques Used:

    ASC is involved in IL-1β production
    Figure Legend Snippet: ASC is involved in IL-1β production

    Techniques Used:

    23) Product Images from "Influenza A Virus as a Predisposing Factor for Cryptococcosis"

    Article Title: Influenza A Virus as a Predisposing Factor for Cryptococcosis

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00419

    Measurement of cytokines and chemokine in the lungs and myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) activities in the lungs of mice after 3 days of influenza A virus (IAV) infection compared to non-infected (NI) group. Six mice per group were infected only with IAV or non-infected (NI) to investigate the immunological response induced by influenza. The animals were ethically euthanized 3 days after IAV infection to assess cytokines and chemokine levels and MPO and NAG activities in the lungs. Fold increase of mRNA of IFN-α4 (A ) and IFN-β (B) . Levels of IFN-γ (C) , TNF-α (D) , CXCL-1 (E) , IL-1β (F) , IL-6 (G) , IL-4 (H) , and IL-10 (I) . The relative number of neutrophils was indirectly measured by the MPO activity (J) and the relative number macrophages was indirectly measured by the NAG activity (K) . * p
    Figure Legend Snippet: Measurement of cytokines and chemokine in the lungs and myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) activities in the lungs of mice after 3 days of influenza A virus (IAV) infection compared to non-infected (NI) group. Six mice per group were infected only with IAV or non-infected (NI) to investigate the immunological response induced by influenza. The animals were ethically euthanized 3 days after IAV infection to assess cytokines and chemokine levels and MPO and NAG activities in the lungs. Fold increase of mRNA of IFN-α4 (A ) and IFN-β (B) . Levels of IFN-γ (C) , TNF-α (D) , CXCL-1 (E) , IL-1β (F) , IL-6 (G) , IL-4 (H) , and IL-10 (I) . The relative number of neutrophils was indirectly measured by the MPO activity (J) and the relative number macrophages was indirectly measured by the NAG activity (K) . * p

    Techniques Used: Mouse Assay, Infection, Activity Assay

    24) Product Images from "The IL23R R381Q Gene Variant Protects against Immune-Mediated Diseases by Impairing IL-23-Induced Th17 Effector Response in Humans"

    Article Title: The IL23R R381Q Gene Variant Protects against Immune-Mediated Diseases by Impairing IL-23-Induced Th17 Effector Response in Humans

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0017160

    IL-1β/IL-23-polarized cells express Th17 markers and produce IL-17A. ( A ) Naive CD4 + T cells were cultured for 6 d in the presence of anti-CD3/CD28 coated beads, with or without IL-1β. On d6 cells were counted and IL-2 added to each condition, together with IL-1β and IL-23, where required. IL-1β/IL-23-polarized Th17 cells were collected after 13 d of culture for phenotypic analysis or rested and assayed for Th17 cytokine on d14/15. ( B ) mRNA expression levels of RORC, IL-23R and CCR6 on d13 were significantly increased in IL-1β/Il-23-polarized cells. ( C ) IL-17A production on d15 was significantly enhanced in IL-1β/Il-23-polarized cells. Results are representative of 5 experiments. Each symbol corresponds to one individual, horizontal bars represent medians. Wilcoxon signed rank test was performed. ** P
    Figure Legend Snippet: IL-1β/IL-23-polarized cells express Th17 markers and produce IL-17A. ( A ) Naive CD4 + T cells were cultured for 6 d in the presence of anti-CD3/CD28 coated beads, with or without IL-1β. On d6 cells were counted and IL-2 added to each condition, together with IL-1β and IL-23, where required. IL-1β/IL-23-polarized Th17 cells were collected after 13 d of culture for phenotypic analysis or rested and assayed for Th17 cytokine on d14/15. ( B ) mRNA expression levels of RORC, IL-23R and CCR6 on d13 were significantly increased in IL-1β/Il-23-polarized cells. ( C ) IL-17A production on d15 was significantly enhanced in IL-1β/Il-23-polarized cells. Results are representative of 5 experiments. Each symbol corresponds to one individual, horizontal bars represent medians. Wilcoxon signed rank test was performed. ** P

    Techniques Used: Cell Culture, Expressing

    IL23R R381Q gene variant reduces IL-23-induced IL-17A production and STAT3 phosphorylation. IL-1β-polarized Th17 cells collected on d13 were stimulated with IL-23. ( A ) IL-17A production in response to 48 h IL-23 stimulation was significantly reduced in IL-1β-polarized Th17 cells from A (red dots) compared to G (green dots) group. Data are expressed as net IL-17A production in response to IL-23. ( B ) IL-17A mRNA expression in response to 24 h IL-23 stimulation was significantly reduced in IL-1β-polarized Th17 cells from A compared to G group. Data are expressed as fold increase compared to unstimulated cells. ( C ) Representative flow cytometry histograms from G (green frame) and A (red frame) donors showing STAT-3 phosphorylation in response to 15 min IL-23-stimulation (blue line) as compared to unstimulated control cells (black line) and expressed as fold Median Fluorescence Intensity (MFI). ( D ) STAT3 phosphorylation in response to IL-23-stimulation was reduced in IL-1β-polarized cells from A compared to G group. Each symbol represents one individual donor; horizontal bars represent medians (A) or means (B, D). Mann-Whitney (A) or unpaired t test (B, D) was performed, *P
    Figure Legend Snippet: IL23R R381Q gene variant reduces IL-23-induced IL-17A production and STAT3 phosphorylation. IL-1β-polarized Th17 cells collected on d13 were stimulated with IL-23. ( A ) IL-17A production in response to 48 h IL-23 stimulation was significantly reduced in IL-1β-polarized Th17 cells from A (red dots) compared to G (green dots) group. Data are expressed as net IL-17A production in response to IL-23. ( B ) IL-17A mRNA expression in response to 24 h IL-23 stimulation was significantly reduced in IL-1β-polarized Th17 cells from A compared to G group. Data are expressed as fold increase compared to unstimulated cells. ( C ) Representative flow cytometry histograms from G (green frame) and A (red frame) donors showing STAT-3 phosphorylation in response to 15 min IL-23-stimulation (blue line) as compared to unstimulated control cells (black line) and expressed as fold Median Fluorescence Intensity (MFI). ( D ) STAT3 phosphorylation in response to IL-23-stimulation was reduced in IL-1β-polarized cells from A compared to G group. Each symbol represents one individual donor; horizontal bars represent medians (A) or means (B, D). Mann-Whitney (A) or unpaired t test (B, D) was performed, *P

    Techniques Used: Variant Assay, Expressing, Flow Cytometry, Cytometry, Fluorescence, MANN-WHITNEY

    IL-1β-polarized cells express Th17 markers and produce IL-17A in response to IL-23. ( A ) Naïve CD4 + T cells were cultured for 6 d in the presence of anti-CD3/CD28 coated-beads, with or without IL-1β. On d6 cells were counted and IL-2 added to each condition, together with IL-1β, where required. IL-1β-polarized Th17 cells were collected after 13 d of culture for phenotypic analysis or assayed for Th17 cytokine on d14/15. ( B ) mRNA expression levels of RORC, IL-23R and CCR6 on d13 were significantly increased in IL-1β-polarized cells. ( C ) IL-17A production in response to PMA/Ionomycin (PMA) was significantly enhanced in IL-1β-polarized cells. ( D ) IL-1β-polarized cells produced higher level of IL-17A in response to IL-23 than control cells. Results are representative of 5 experiments. Each symbol corresponds to one individual, horizontal bars represent medians. Wilcoxon signed rank test was performed. *P
    Figure Legend Snippet: IL-1β-polarized cells express Th17 markers and produce IL-17A in response to IL-23. ( A ) Naïve CD4 + T cells were cultured for 6 d in the presence of anti-CD3/CD28 coated-beads, with or without IL-1β. On d6 cells were counted and IL-2 added to each condition, together with IL-1β, where required. IL-1β-polarized Th17 cells were collected after 13 d of culture for phenotypic analysis or assayed for Th17 cytokine on d14/15. ( B ) mRNA expression levels of RORC, IL-23R and CCR6 on d13 were significantly increased in IL-1β-polarized cells. ( C ) IL-17A production in response to PMA/Ionomycin (PMA) was significantly enhanced in IL-1β-polarized cells. ( D ) IL-1β-polarized cells produced higher level of IL-17A in response to IL-23 than control cells. Results are representative of 5 experiments. Each symbol corresponds to one individual, horizontal bars represent medians. Wilcoxon signed rank test was performed. *P

    Techniques Used: Cell Culture, Expressing, Produced

    IL23R R381Q gene variant and IL-1β-polarized Th17 cells. IL-1β-polarized Th17 cells were used for phenotypic analysis on d13. ( A ) Percentage of CCR6 + IL-23R + IL-1β-polarized cells did not significantly differ between G (green triangles) and A group (red triangles). ( B ) mRNA expression levels of RORC, IL-23R and CCR6 did not differ between G and A group. Each symbol represents one individual donor; horizontal bars represent medians (A) or means (B). Mann Whitney test (A) or unpaired t test (B) were performed yielding P values > 0.05 for all panels.
    Figure Legend Snippet: IL23R R381Q gene variant and IL-1β-polarized Th17 cells. IL-1β-polarized Th17 cells were used for phenotypic analysis on d13. ( A ) Percentage of CCR6 + IL-23R + IL-1β-polarized cells did not significantly differ between G (green triangles) and A group (red triangles). ( B ) mRNA expression levels of RORC, IL-23R and CCR6 did not differ between G and A group. Each symbol represents one individual donor; horizontal bars represent medians (A) or means (B). Mann Whitney test (A) or unpaired t test (B) were performed yielding P values > 0.05 for all panels.

    Techniques Used: Variant Assay, Expressing, MANN-WHITNEY

    IL23R R381Q gene variant does not affect Th17 differentiation. IL-1β/IL-23-polarized Th17 cells were used for phenotypic analysis (on d13) or assayed for IL-17A mRNA expression (on d14) or secretion (on d15). ( A ) Percentage of CCR6 + IL-23R + IL-1β/IL-23-polarized Th17 cells did not significantly differ between G (green triangles) and A group (red triangles). ( B ) mRNA expression levels of RORC, IL-23R and CCR6 did not differ between G and A group. ( C ) IL-17A production and ( D ) IL-17A mRNA expression did not differ between G and A group. Each symbol represents one individual, horizontal bars represent medians. Mann Whitney test was performed yielding P values > 0.05 for all panels.
    Figure Legend Snippet: IL23R R381Q gene variant does not affect Th17 differentiation. IL-1β/IL-23-polarized Th17 cells were used for phenotypic analysis (on d13) or assayed for IL-17A mRNA expression (on d14) or secretion (on d15). ( A ) Percentage of CCR6 + IL-23R + IL-1β/IL-23-polarized Th17 cells did not significantly differ between G (green triangles) and A group (red triangles). ( B ) mRNA expression levels of RORC, IL-23R and CCR6 did not differ between G and A group. ( C ) IL-17A production and ( D ) IL-17A mRNA expression did not differ between G and A group. Each symbol represents one individual, horizontal bars represent medians. Mann Whitney test was performed yielding P values > 0.05 for all panels.

    Techniques Used: Variant Assay, Expressing, MANN-WHITNEY

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    Article Snippet: .. In separate experimental groups, NANT cells were cultured in the presence or absence of exogenous human recombinant IL-5 (100 ng ml−1 ; R & D Systems, Minneapolis, MN, U.S.A.), or anti-IL-5Rα monoclonal neutralizing antibody (2.5 μg ml−1 ; Catalogue number AF-253-NA, R & D Systems, Minneapolis, MN, U.S.A.), or dexamethasone (10−6 M ). .. In the studies with dexamethasone, NANT cells were incubated with either dexamethasone or with diluent alone, the latter acting as time control, for 16 h. To determine whether any residual T lymphocytes in NANT cell preparation could act as a source of IL-5, NANT cells cultured from 14 asthmatics and 14 normal subjects were compared with those co-cultured with autologous T lymphocytes (at 5×104 cells ml−1 ) from the same study subjects.

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    Article Title: Interleukin-5 in growth and differentiation of blood eosinophil progenitors in asthma: effect of glucocorticoids
    Article Snippet: .. In separate experimental groups, NANT cells were cultured in the presence or absence of exogenous human recombinant IL-5 (100 ng ml−1 ; R & D Systems, Minneapolis, MN, U.S.A.), or anti-IL-5Rα monoclonal neutralizing antibody (2.5 μg ml−1 ; Catalogue number AF-253-NA, R & D Systems, Minneapolis, MN, U.S.A.), or dexamethasone (10−6 M ). .. In the studies with dexamethasone, NANT cells were incubated with either dexamethasone or with diluent alone, the latter acting as time control, for 16 h. To determine whether any residual T lymphocytes in NANT cell preparation could act as a source of IL-5, NANT cells cultured from 14 asthmatics and 14 normal subjects were compared with those co-cultured with autologous T lymphocytes (at 5×104 cells ml−1 ) from the same study subjects.

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    R&D Systems rat il 1β
    <t>IL-1β</t> signaling in the CNS produces muscle catabolism. (a) Atrophy gene expression in mouse Gn muscle (Gn) after acute i.c.v. IL-1β injection (10 ng, n = 6–9/group) or Veh. (b) Akt and p38 phosphorylation in Gn after acute i.c.v. IL-1β was measured by Western blotting ( n = 6–9/group). (c–g) Rats were i.c.v. infused for 3 d with IL-1β (10 ng/h) or Veh ( n = 8–11/group). A subset of Veh-treated animals were PF to IL-1β–treated animals. (c) Atrophy gene expression in rat muscle (Gn, Soleus, and EDL) after 3-d i.c.v. IL-1β or Veh infusion was measured by real-time PCR. Reported values are relative to GAPDH. (d) Body composition after 3-d IL-1β infusion. (e) Muscle weight normalized to initial body weight (BW) and weight change relative to Veh. (f) Mean fiber CSA in WGn and soleus and frequency distribution in Gn. (g) WGn immunostained for laminin (red), delineating fiber area, and Myosin IIb (green). Bars, 100 µm. Data are represented as mean ± SEM. *, P
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    R&D Systems antibody against il 1β
    MiR-27a-3p inhibits OB progression through the EMT pathway. a Smad3 transcription was significantly promoted by adding the miR-27a-3p inhibitor in mouse airway epithelial cells. b The influence of miR-27a-3p on protein expression levels of p-Smad3, Smad3, and EMT markers in mouse airway epithelial cells was examined using Western blot assay. c Adding the miR-27a-3p inhibitor significantly promoted the <t>IL-1β</t> content in the medium of mouse airway epithelial cells. d Transfection of miR-27a-3p mimic significantly inhibited Smad3 transcription in NIH-3T3 and LLC cells. e Influence of miR-27a-3p on the protein expression levels of p-Smad3, Smad3, and EMT markers in NIH-3T3 cells was examined using Western blot assay. f Transfection of miR-27a-3p mimic significantly inhibited the IL-1β content in the medium of NIH-3T3 and LLC cells. g The influence of miR-27a-3p on protein expression levels of p-Smad3, Smad3, and EMT markers in LLC cells was examined using Western blot assay (mean ± SD; n = 3 in triplicate; ** P
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    Dioscin attenuated ECM degradation in <t>IL-1β-stimulated</t> human NP cells. ( A, B ) Relative mRNA level of ( A ) collagen II and ( B ) aggrecan were examined by PCR. ( C ) Fluorescence immunostaining of collagen II protein (green) and nucleus (blue). The results are presented as the means ±SD. * P
    Il 1β, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    <t>IL1β-IR</t> in the Sctx at 1100 (light) or 2300 (dark) IL1β-IR cells increased in Sctx layers II–V at 2300 h (C, D F) in comparison to 1100 h (A, B, E). The IL1β-IR appears in the cytoplasm and extends into the apical dendrite of the large pyramidal neurons in layer V at 2300 h (D F). Bar = 0.05 mm for A–D; Bar =0.025 mm for E F.
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    Image Search Results


    IL-1β signaling in the CNS produces muscle catabolism. (a) Atrophy gene expression in mouse Gn muscle (Gn) after acute i.c.v. IL-1β injection (10 ng, n = 6–9/group) or Veh. (b) Akt and p38 phosphorylation in Gn after acute i.c.v. IL-1β was measured by Western blotting ( n = 6–9/group). (c–g) Rats were i.c.v. infused for 3 d with IL-1β (10 ng/h) or Veh ( n = 8–11/group). A subset of Veh-treated animals were PF to IL-1β–treated animals. (c) Atrophy gene expression in rat muscle (Gn, Soleus, and EDL) after 3-d i.c.v. IL-1β or Veh infusion was measured by real-time PCR. Reported values are relative to GAPDH. (d) Body composition after 3-d IL-1β infusion. (e) Muscle weight normalized to initial body weight (BW) and weight change relative to Veh. (f) Mean fiber CSA in WGn and soleus and frequency distribution in Gn. (g) WGn immunostained for laminin (red), delineating fiber area, and Myosin IIb (green). Bars, 100 µm. Data are represented as mean ± SEM. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Central nervous system inflammation induces muscle atrophy via activation of the hypothalamic-pituitary-adrenal axis

    doi: 10.1084/jem.20111020

    Figure Lengend Snippet: IL-1β signaling in the CNS produces muscle catabolism. (a) Atrophy gene expression in mouse Gn muscle (Gn) after acute i.c.v. IL-1β injection (10 ng, n = 6–9/group) or Veh. (b) Akt and p38 phosphorylation in Gn after acute i.c.v. IL-1β was measured by Western blotting ( n = 6–9/group). (c–g) Rats were i.c.v. infused for 3 d with IL-1β (10 ng/h) or Veh ( n = 8–11/group). A subset of Veh-treated animals were PF to IL-1β–treated animals. (c) Atrophy gene expression in rat muscle (Gn, Soleus, and EDL) after 3-d i.c.v. IL-1β or Veh infusion was measured by real-time PCR. Reported values are relative to GAPDH. (d) Body composition after 3-d IL-1β infusion. (e) Muscle weight normalized to initial body weight (BW) and weight change relative to Veh. (f) Mean fiber CSA in WGn and soleus and frequency distribution in Gn. (g) WGn immunostained for laminin (red), delineating fiber area, and Myosin IIb (green). Bars, 100 µm. Data are represented as mean ± SEM. *, P

    Article Snippet: Cannulas were implanted into the lateral ventricle under isoflurane anesthesia, using a stereotactic alignment instrument (Kopf) at the following coordinates relative to bregma: −1.5 mm X, −1.0 mm Y, and −4.0 mm Z. Mini osmotic pumps delivering 10 ng rat IL-1β per hour (R & D Systems) were connected via polyethylene catheters to the cannulas and implanted subcutaneously.

    Techniques: Expressing, Injection, Western Blot, Real-time Polymerase Chain Reaction

    Glucocorticoid-driven gene expression signatures are present in skeletal muscle during central and peripheral inflammation. (a) Gene expression in ADX mouse Gn muscle 8 h after acute i.c.v. 10-ng IL-1β injection or Veh was measured by real-time PCR ( n = 6–8/group). (b) Muscle gene expression in mouse Gn 8 h after i.p. LPS (250 µg/kg) or Veh was measured by real-time PCR ( n = 5–7/group). (c) Gene expression in Gn in mice bearing LLC tumors (or sham) was measured by real-time PCR ( n = 5–9/group). Reported values are relative to GAPDH. Data represented as mean ± SEM. The dotted line in a represents Veh-treated controls at a relative quantity of 1. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Central nervous system inflammation induces muscle atrophy via activation of the hypothalamic-pituitary-adrenal axis

    doi: 10.1084/jem.20111020

    Figure Lengend Snippet: Glucocorticoid-driven gene expression signatures are present in skeletal muscle during central and peripheral inflammation. (a) Gene expression in ADX mouse Gn muscle 8 h after acute i.c.v. 10-ng IL-1β injection or Veh was measured by real-time PCR ( n = 6–8/group). (b) Muscle gene expression in mouse Gn 8 h after i.p. LPS (250 µg/kg) or Veh was measured by real-time PCR ( n = 5–7/group). (c) Gene expression in Gn in mice bearing LLC tumors (or sham) was measured by real-time PCR ( n = 5–9/group). Reported values are relative to GAPDH. Data represented as mean ± SEM. The dotted line in a represents Veh-treated controls at a relative quantity of 1. *, P

    Article Snippet: Cannulas were implanted into the lateral ventricle under isoflurane anesthesia, using a stereotactic alignment instrument (Kopf) at the following coordinates relative to bregma: −1.5 mm X, −1.0 mm Y, and −4.0 mm Z. Mini osmotic pumps delivering 10 ng rat IL-1β per hour (R & D Systems) were connected via polyethylene catheters to the cannulas and implanted subcutaneously.

    Techniques: Expressing, Injection, Real-time Polymerase Chain Reaction, Mouse Assay

    Central IL-1β treatment induces rapid and dynamic changes in skeletal muscle gene expression. WT mice received lateral ventricle injections of IL-1β (I) or Veh (V). Food was removed from the cages at the time of injection, and animals were sacrificed at 2, 4, and 8 h after the injection ( n = 3–5/group). Skeletal muscle RNA was hybridized to Affymetrix Exon 1.0 ST arrays. Data were analyzed for a false discovery rate of 0.01. Genes that were > 1.75-fold up- or down-regulated at any time point were categorized based on their annotated and published functions. Heat map values are log 2 normalized array intensities. Genes with differential expression confirmed by real-time PCR are indicated in red ( Fig. S1 ). § indicates genes that are annotated as glucocorticoid responsive or are regulated in muscle by glucocorticoids in published array datasets.

    Journal: The Journal of Experimental Medicine

    Article Title: Central nervous system inflammation induces muscle atrophy via activation of the hypothalamic-pituitary-adrenal axis

    doi: 10.1084/jem.20111020

    Figure Lengend Snippet: Central IL-1β treatment induces rapid and dynamic changes in skeletal muscle gene expression. WT mice received lateral ventricle injections of IL-1β (I) or Veh (V). Food was removed from the cages at the time of injection, and animals were sacrificed at 2, 4, and 8 h after the injection ( n = 3–5/group). Skeletal muscle RNA was hybridized to Affymetrix Exon 1.0 ST arrays. Data were analyzed for a false discovery rate of 0.01. Genes that were > 1.75-fold up- or down-regulated at any time point were categorized based on their annotated and published functions. Heat map values are log 2 normalized array intensities. Genes with differential expression confirmed by real-time PCR are indicated in red ( Fig. S1 ). § indicates genes that are annotated as glucocorticoid responsive or are regulated in muscle by glucocorticoids in published array datasets.

    Article Snippet: Cannulas were implanted into the lateral ventricle under isoflurane anesthesia, using a stereotactic alignment instrument (Kopf) at the following coordinates relative to bregma: −1.5 mm X, −1.0 mm Y, and −4.0 mm Z. Mini osmotic pumps delivering 10 ng rat IL-1β per hour (R & D Systems) were connected via polyethylene catheters to the cannulas and implanted subcutaneously.

    Techniques: Expressing, Mouse Assay, Injection, Real-time Polymerase Chain Reaction

    Peripheral infusion of low dose IL-1β does not cause atrophy. Rats were i.p. infused (10 ng/h, n = 7–8/group) for 3 d with IL-1β or Veh solution. (a) Atrophy gene expression in rat muscle (Gn, Soleus, and EDL) after 3-d i.p. IL-1β infusion was measured by real-time PCR. Reported values are relative to GAPDH. (b) Muscle weight normalized to initial body weight (BW). (c) Body composition after 3-d IL-1β infusion. (d) Mean fiber CSA in WGn and soleus and frequency distribution in Gn. (e) WGn immunostained for laminin (green) delineating fiber area. Bars, 100 µm. Data are represented as mean ± SEM.

    Journal: The Journal of Experimental Medicine

    Article Title: Central nervous system inflammation induces muscle atrophy via activation of the hypothalamic-pituitary-adrenal axis

    doi: 10.1084/jem.20111020

    Figure Lengend Snippet: Peripheral infusion of low dose IL-1β does not cause atrophy. Rats were i.p. infused (10 ng/h, n = 7–8/group) for 3 d with IL-1β or Veh solution. (a) Atrophy gene expression in rat muscle (Gn, Soleus, and EDL) after 3-d i.p. IL-1β infusion was measured by real-time PCR. Reported values are relative to GAPDH. (b) Muscle weight normalized to initial body weight (BW). (c) Body composition after 3-d IL-1β infusion. (d) Mean fiber CSA in WGn and soleus and frequency distribution in Gn. (e) WGn immunostained for laminin (green) delineating fiber area. Bars, 100 µm. Data are represented as mean ± SEM.

    Article Snippet: Cannulas were implanted into the lateral ventricle under isoflurane anesthesia, using a stereotactic alignment instrument (Kopf) at the following coordinates relative to bregma: −1.5 mm X, −1.0 mm Y, and −4.0 mm Z. Mini osmotic pumps delivering 10 ng rat IL-1β per hour (R & D Systems) were connected via polyethylene catheters to the cannulas and implanted subcutaneously.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Signaling through the MC4R is not a major contributor to the muscle catabolism independent of food intake. (a and b) Atrophy gene expression in WT and MC4RKO mouse Gn 8 h after acute i.c.v. IL-1β injection (10 ng, n = 8–9/group; a) and in rat Gn 6 h after acute i.c.v. MTII (1 nmol, n = 6–9/group; b) or Veh injection was measured by real-time PCR. (c–e) Rats were treated for 36 h (chronic) with MTII or Veh (1 nmol/12 h × 4, n = 7–8/group). Veh-treated animals were PF to the MTII treatment group. (c) Gene expression in rat Gn and soleus after chronic MTII treatment was measured by real-time PCR. Reported values are relative to GAPDH . (d) Body composition after chronic MTII treatment. (e) Muscle weight normalized to initial body weight (BW). The dotted line in a represents Veh-treated controls at a relative quantity of 1. Data are represented as mean ± SEM. **, P

    Journal: The Journal of Experimental Medicine

    Article Title: Central nervous system inflammation induces muscle atrophy via activation of the hypothalamic-pituitary-adrenal axis

    doi: 10.1084/jem.20111020

    Figure Lengend Snippet: Signaling through the MC4R is not a major contributor to the muscle catabolism independent of food intake. (a and b) Atrophy gene expression in WT and MC4RKO mouse Gn 8 h after acute i.c.v. IL-1β injection (10 ng, n = 8–9/group; a) and in rat Gn 6 h after acute i.c.v. MTII (1 nmol, n = 6–9/group; b) or Veh injection was measured by real-time PCR. (c–e) Rats were treated for 36 h (chronic) with MTII or Veh (1 nmol/12 h × 4, n = 7–8/group). Veh-treated animals were PF to the MTII treatment group. (c) Gene expression in rat Gn and soleus after chronic MTII treatment was measured by real-time PCR. Reported values are relative to GAPDH . (d) Body composition after chronic MTII treatment. (e) Muscle weight normalized to initial body weight (BW). The dotted line in a represents Veh-treated controls at a relative quantity of 1. Data are represented as mean ± SEM. **, P

    Article Snippet: Cannulas were implanted into the lateral ventricle under isoflurane anesthesia, using a stereotactic alignment instrument (Kopf) at the following coordinates relative to bregma: −1.5 mm X, −1.0 mm Y, and −4.0 mm Z. Mini osmotic pumps delivering 10 ng rat IL-1β per hour (R & D Systems) were connected via polyethylene catheters to the cannulas and implanted subcutaneously.

    Techniques: Expressing, Injection, Real-time Polymerase Chain Reaction

    CNS inflammation and muscle catabolism occur simultaneously. (a) Atrophy gene expression in mouse Gn and cytokine expression in mouse hypothalamic blocks (Hypo) 8 h after i.p. LPS (250 µg/kg) or Veh was measured by real-time PCR ( n = 5–7/group for Gn and 5/group for Hypo). Values are relative to GAPDH for Gn and 18s RNA for Hypo. (b) In situ hybridization for IL-1β mRNA in the arcuate nucleus of the hypothalamus (−2.80 relative to bregma; 3V, third ventricle) 8 h after LPS administration. Veh-treated animals showed no specific signal at this exposure time. (c) Atrophy gene expression in Gn and cytokine expression in hypothalamic blocks in mice bearing LLC tumors (or sham) were measured by real-time PCR ( n = 5–9/group). Values are relative to GAPDH for Gn and 18s RNA for Hypo. (d) Gn muscle weight normalized to initial body weight (BW) and muscle weight change relative to Veh in LLC tumor bearing animals ( n = 5–9/group). Bar, 100 µm. Data are represented as mean ± SEM. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Central nervous system inflammation induces muscle atrophy via activation of the hypothalamic-pituitary-adrenal axis

    doi: 10.1084/jem.20111020

    Figure Lengend Snippet: CNS inflammation and muscle catabolism occur simultaneously. (a) Atrophy gene expression in mouse Gn and cytokine expression in mouse hypothalamic blocks (Hypo) 8 h after i.p. LPS (250 µg/kg) or Veh was measured by real-time PCR ( n = 5–7/group for Gn and 5/group for Hypo). Values are relative to GAPDH for Gn and 18s RNA for Hypo. (b) In situ hybridization for IL-1β mRNA in the arcuate nucleus of the hypothalamus (−2.80 relative to bregma; 3V, third ventricle) 8 h after LPS administration. Veh-treated animals showed no specific signal at this exposure time. (c) Atrophy gene expression in Gn and cytokine expression in hypothalamic blocks in mice bearing LLC tumors (or sham) were measured by real-time PCR ( n = 5–9/group). Values are relative to GAPDH for Gn and 18s RNA for Hypo. (d) Gn muscle weight normalized to initial body weight (BW) and muscle weight change relative to Veh in LLC tumor bearing animals ( n = 5–9/group). Bar, 100 µm. Data are represented as mean ± SEM. *, P

    Article Snippet: Cannulas were implanted into the lateral ventricle under isoflurane anesthesia, using a stereotactic alignment instrument (Kopf) at the following coordinates relative to bregma: −1.5 mm X, −1.0 mm Y, and −4.0 mm Z. Mini osmotic pumps delivering 10 ng rat IL-1β per hour (R & D Systems) were connected via polyethylene catheters to the cannulas and implanted subcutaneously.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, In Situ Hybridization, Mouse Assay

    HPA axis activation is necessary for i.c.v. IL-1β–induced muscle catabolism. (a and b) Muscle atrophy gene expression in mifepristone (Mife) or Veh-treated ( n = 6–8/group; a) and ADX or sham ( n = 7–8/group; b) mouse Gn 8 h after acute i.c.v. 10-ng IL-1β injection was measured by real-time PCR. (c–g) ADX rats were treated for 3 d with chronic i.c.v. IL-1β or Veh (10 ng/h, n = 8–9/group). Veh-treated animals were PF to the IL-1β–treated group. (c) Atrophy gene expression in ADX rat muscle after 3-d chronic i.c.v. IL-1β infusion was measured by real-time PCR. Reported values are relative to GAPDH. (d) Body composition. (e) Muscle weight normalized to initial body weight (BW). (f) Mean fiber CSA in WGn and soleus and frequency distribution in WGn. (g) WGn immunostained for laminin (green) delineating fiber area. The dotted lines in a and b represent Veh-treated controls at a relative quantity of 1. Bars, 100 µm. Data are represented as mean ± SEM. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Central nervous system inflammation induces muscle atrophy via activation of the hypothalamic-pituitary-adrenal axis

    doi: 10.1084/jem.20111020

    Figure Lengend Snippet: HPA axis activation is necessary for i.c.v. IL-1β–induced muscle catabolism. (a and b) Muscle atrophy gene expression in mifepristone (Mife) or Veh-treated ( n = 6–8/group; a) and ADX or sham ( n = 7–8/group; b) mouse Gn 8 h after acute i.c.v. 10-ng IL-1β injection was measured by real-time PCR. (c–g) ADX rats were treated for 3 d with chronic i.c.v. IL-1β or Veh (10 ng/h, n = 8–9/group). Veh-treated animals were PF to the IL-1β–treated group. (c) Atrophy gene expression in ADX rat muscle after 3-d chronic i.c.v. IL-1β infusion was measured by real-time PCR. Reported values are relative to GAPDH. (d) Body composition. (e) Muscle weight normalized to initial body weight (BW). (f) Mean fiber CSA in WGn and soleus and frequency distribution in WGn. (g) WGn immunostained for laminin (green) delineating fiber area. The dotted lines in a and b represent Veh-treated controls at a relative quantity of 1. Bars, 100 µm. Data are represented as mean ± SEM. *, P

    Article Snippet: Cannulas were implanted into the lateral ventricle under isoflurane anesthesia, using a stereotactic alignment instrument (Kopf) at the following coordinates relative to bregma: −1.5 mm X, −1.0 mm Y, and −4.0 mm Z. Mini osmotic pumps delivering 10 ng rat IL-1β per hour (R & D Systems) were connected via polyethylene catheters to the cannulas and implanted subcutaneously.

    Techniques: Activation Assay, Expressing, Injection, Real-time Polymerase Chain Reaction

    MiR-27a-3p inhibits OB progression through the EMT pathway. a Smad3 transcription was significantly promoted by adding the miR-27a-3p inhibitor in mouse airway epithelial cells. b The influence of miR-27a-3p on protein expression levels of p-Smad3, Smad3, and EMT markers in mouse airway epithelial cells was examined using Western blot assay. c Adding the miR-27a-3p inhibitor significantly promoted the IL-1β content in the medium of mouse airway epithelial cells. d Transfection of miR-27a-3p mimic significantly inhibited Smad3 transcription in NIH-3T3 and LLC cells. e Influence of miR-27a-3p on the protein expression levels of p-Smad3, Smad3, and EMT markers in NIH-3T3 cells was examined using Western blot assay. f Transfection of miR-27a-3p mimic significantly inhibited the IL-1β content in the medium of NIH-3T3 and LLC cells. g The influence of miR-27a-3p on protein expression levels of p-Smad3, Smad3, and EMT markers in LLC cells was examined using Western blot assay (mean ± SD; n = 3 in triplicate; ** P

    Journal: Cell Stress & Chaperones

    Article Title: MiR-27a-3p downregulation contributes to the development of occlusive bronchiolitis

    doi: 10.1007/s12192-019-01026-7

    Figure Lengend Snippet: MiR-27a-3p inhibits OB progression through the EMT pathway. a Smad3 transcription was significantly promoted by adding the miR-27a-3p inhibitor in mouse airway epithelial cells. b The influence of miR-27a-3p on protein expression levels of p-Smad3, Smad3, and EMT markers in mouse airway epithelial cells was examined using Western blot assay. c Adding the miR-27a-3p inhibitor significantly promoted the IL-1β content in the medium of mouse airway epithelial cells. d Transfection of miR-27a-3p mimic significantly inhibited Smad3 transcription in NIH-3T3 and LLC cells. e Influence of miR-27a-3p on the protein expression levels of p-Smad3, Smad3, and EMT markers in NIH-3T3 cells was examined using Western blot assay. f Transfection of miR-27a-3p mimic significantly inhibited the IL-1β content in the medium of NIH-3T3 and LLC cells. g The influence of miR-27a-3p on protein expression levels of p-Smad3, Smad3, and EMT markers in LLC cells was examined using Western blot assay (mean ± SD; n = 3 in triplicate; ** P

    Article Snippet: The interleukin (IL)-1β levels in the cell culture medium with different treatments were quantified using sand with ELISA kit, which comprised 96-well microplates coated with an antibody against IL-1β (R & D Systems, USA).

    Techniques: Expressing, Western Blot, Transfection

    Dioscin attenuated ECM degradation in IL-1β-stimulated human NP cells. ( A, B ) Relative mRNA level of ( A ) collagen II and ( B ) aggrecan were examined by PCR. ( C ) Fluorescence immunostaining of collagen II protein (green) and nucleus (blue). The results are presented as the means ±SD. * P

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

    Article Title: Dioscin Attenuates Interleukin 1β (IL-1β)-Induced Catabolism and Apoptosis via Modulating the Toll-Like Receptor 4 (TLR4)/Nuclear Factor kappa B (NF-κB) Signaling in Human Nucleus Pulposus Cells

    doi: 10.12659/MSM.923386

    Figure Lengend Snippet: Dioscin attenuated ECM degradation in IL-1β-stimulated human NP cells. ( A, B ) Relative mRNA level of ( A ) collagen II and ( B ) aggrecan were examined by PCR. ( C ) Fluorescence immunostaining of collagen II protein (green) and nucleus (blue). The results are presented as the means ±SD. * P

    Article Snippet: IL-1β was acquired from R & D Systems (St. Paul, MN, USA).

    Techniques: Polymerase Chain Reaction, Fluorescence, Immunostaining

    Effects of dioscin on IL-1β-activated cell viability in human NP cells. ( A ) The cytotoxic effects of dioscin on human NP cells were examined at different concentrations by the CCK-8 method. ( B ) CCK-8 analysis of dioscin-treated human NP cells stimulated by IL-1β. The results are presented as the means ±SD. * P

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

    Article Title: Dioscin Attenuates Interleukin 1β (IL-1β)-Induced Catabolism and Apoptosis via Modulating the Toll-Like Receptor 4 (TLR4)/Nuclear Factor kappa B (NF-κB) Signaling in Human Nucleus Pulposus Cells

    doi: 10.12659/MSM.923386

    Figure Lengend Snippet: Effects of dioscin on IL-1β-activated cell viability in human NP cells. ( A ) The cytotoxic effects of dioscin on human NP cells were examined at different concentrations by the CCK-8 method. ( B ) CCK-8 analysis of dioscin-treated human NP cells stimulated by IL-1β. The results are presented as the means ±SD. * P

    Article Snippet: IL-1β was acquired from R & D Systems (St. Paul, MN, USA).

    Techniques: CCK-8 Assay

    Effect of dioscin on IL-1β–activated apoptotic activity in human NP cells. ( A, B ) Apoptotic activity was evaluated using the TUNEL method. ( C–F ) Western blotting and quantitative analysis of cleaved caspase-3, -9, and Bcl-2 expressions. ( G ) Fluorescence immunostaining of cleaved caspase-3 (red) and nucleus (blue). The results are presented as the means ±SD. * P

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

    Article Title: Dioscin Attenuates Interleukin 1β (IL-1β)-Induced Catabolism and Apoptosis via Modulating the Toll-Like Receptor 4 (TLR4)/Nuclear Factor kappa B (NF-κB) Signaling in Human Nucleus Pulposus Cells

    doi: 10.12659/MSM.923386

    Figure Lengend Snippet: Effect of dioscin on IL-1β–activated apoptotic activity in human NP cells. ( A, B ) Apoptotic activity was evaluated using the TUNEL method. ( C–F ) Western blotting and quantitative analysis of cleaved caspase-3, -9, and Bcl-2 expressions. ( G ) Fluorescence immunostaining of cleaved caspase-3 (red) and nucleus (blue). The results are presented as the means ±SD. * P

    Article Snippet: IL-1β was acquired from R & D Systems (St. Paul, MN, USA).

    Techniques: Activity Assay, TUNEL Assay, Western Blot, Fluorescence, Immunostaining

    Dioscin suppressed IL-1β-induced release of proinflammatory factors via regulation of the NF-κB pathway in human NP cells. ( A, B ) Relative mRNA level of ( A ) IL-6 and ( B ) TNF-α were examined by PCR. ( C–F ) Western blotting and quantitative analysis of TLR4, p65, and IκBα. The results are presented as the means ±SD. * P

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

    Article Title: Dioscin Attenuates Interleukin 1β (IL-1β)-Induced Catabolism and Apoptosis via Modulating the Toll-Like Receptor 4 (TLR4)/Nuclear Factor kappa B (NF-κB) Signaling in Human Nucleus Pulposus Cells

    doi: 10.12659/MSM.923386

    Figure Lengend Snippet: Dioscin suppressed IL-1β-induced release of proinflammatory factors via regulation of the NF-κB pathway in human NP cells. ( A, B ) Relative mRNA level of ( A ) IL-6 and ( B ) TNF-α were examined by PCR. ( C–F ) Western blotting and quantitative analysis of TLR4, p65, and IκBα. The results are presented as the means ±SD. * P

    Article Snippet: IL-1β was acquired from R & D Systems (St. Paul, MN, USA).

    Techniques: Polymerase Chain Reaction, Western Blot

    Dioscin suppressed IL-1β-activated catabolic activity in human NP cells. ( A–D ) Relative mRNA level of ( A, B ) ADAMTS4 and ADAMTS5, and ( C, D ) MMP3, MMP13 were measured by PCR. The results are presented as the means ±SD. * P

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

    Article Title: Dioscin Attenuates Interleukin 1β (IL-1β)-Induced Catabolism and Apoptosis via Modulating the Toll-Like Receptor 4 (TLR4)/Nuclear Factor kappa B (NF-κB) Signaling in Human Nucleus Pulposus Cells

    doi: 10.12659/MSM.923386

    Figure Lengend Snippet: Dioscin suppressed IL-1β-activated catabolic activity in human NP cells. ( A–D ) Relative mRNA level of ( A, B ) ADAMTS4 and ADAMTS5, and ( C, D ) MMP3, MMP13 were measured by PCR. The results are presented as the means ±SD. * P

    Article Snippet: IL-1β was acquired from R & D Systems (St. Paul, MN, USA).

    Techniques: Activity Assay, Polymerase Chain Reaction

    IL1β-IR in the Sctx at 1100 (light) or 2300 (dark) IL1β-IR cells increased in Sctx layers II–V at 2300 h (C, D F) in comparison to 1100 h (A, B, E). The IL1β-IR appears in the cytoplasm and extends into the apical dendrite of the large pyramidal neurons in layer V at 2300 h (D F). Bar = 0.05 mm for A–D; Bar =0.025 mm for E F.

    Journal: Brain research

    Article Title: Time of Day differences in the Number of Cytokine-, Neurotrophin- and NeuN-immunoreactive cells in the Rat Somatosensory or Visual Cortex

    doi: 10.1016/j.brainres.2010.04.012

    Figure Lengend Snippet: IL1β-IR in the Sctx at 1100 (light) or 2300 (dark) IL1β-IR cells increased in Sctx layers II–V at 2300 h (C, D F) in comparison to 1100 h (A, B, E). The IL1β-IR appears in the cytoplasm and extends into the apical dendrite of the large pyramidal neurons in layer V at 2300 h (D F). Bar = 0.05 mm for A–D; Bar =0.025 mm for E F.

    Article Snippet: Immunohistochemistry was performed as described previously ( , ), using a polyclonal anti-human Fos antibody produced in rabbit (1:10,000; Calbiochem (Oncogene Research Products), San Diego, CA), goat anti-rat IL1β (0.5 μg/ml, R & D Systems, Inc., Minneapolis, MN), goat anti-rat TNFα (0.5 μg/ml, R & D Systems, Inc., Minneapolis, MN), rabbit anti-mouse NGF that cross reacts with rat NGF (1:2500 or 1:5000; Chemicon, Temecula, CA) or mouse monoclonal antibody to NeuN (1:500; Chemicon, Temecula, CA).

    Techniques:

    IL1β-IR in the Vctx at 1100 (light) or 2300 (dark) IL1β-IR cells increased in Vctx layers V–VI at 2300 h (D F) in comparison to 1100 h (B, E). The IL1β-IR appears in the cytoplasm and extends into the apical dendrite of the large pyramidal neurons in layer V at 2300 h (D F). Bar = 0.05 mm for A–D; Bar =0.025 mm for E F.

    Journal: Brain research

    Article Title: Time of Day differences in the Number of Cytokine-, Neurotrophin- and NeuN-immunoreactive cells in the Rat Somatosensory or Visual Cortex

    doi: 10.1016/j.brainres.2010.04.012

    Figure Lengend Snippet: IL1β-IR in the Vctx at 1100 (light) or 2300 (dark) IL1β-IR cells increased in Vctx layers V–VI at 2300 h (D F) in comparison to 1100 h (B, E). The IL1β-IR appears in the cytoplasm and extends into the apical dendrite of the large pyramidal neurons in layer V at 2300 h (D F). Bar = 0.05 mm for A–D; Bar =0.025 mm for E F.

    Article Snippet: Immunohistochemistry was performed as described previously ( , ), using a polyclonal anti-human Fos antibody produced in rabbit (1:10,000; Calbiochem (Oncogene Research Products), San Diego, CA), goat anti-rat IL1β (0.5 μg/ml, R & D Systems, Inc., Minneapolis, MN), goat anti-rat TNFα (0.5 μg/ml, R & D Systems, Inc., Minneapolis, MN), rabbit anti-mouse NGF that cross reacts with rat NGF (1:2500 or 1:5000; Chemicon, Temecula, CA) or mouse monoclonal antibody to NeuN (1:500; Chemicon, Temecula, CA).

    Techniques: