Journal: Journal of Inflammation Research

Article Title: Electroacupuncture Inhibits the Early Neuroinflammatory Cascade Triggered by TLR2 in the Prodromal Period of PD

doi: 10.2147/JIR.S585729

Figure Lengend Snippet: EA can inhibit the activation of microglia in the pPD stage mediated by TLR2. ( A ) Western blot detection of TLR2 protein expression in the substantia nigra. ( B ) Western blot detection of MyD88 protein expression in the substantia nigra. ( C ) Western blot detection of p-NF-κB-p65 expression in the substantia nigra. ( D ) Western blot detection of NLRP3 protein expression in the substantia nigra. ( E ) Western blot detection of Caspase-1 protein expression in the substantia nigra. ( F ) Western blot detection of GSDMD protein expression in the substantia nigra. ( G ) Western blot detection of IL-1β protein expression in the substantia nigra. (n=6; compared with the control group, *** P <0.001; compared with the model group, ### P <0.001, ## P <0.01, # P <0.05).

Article Snippet: The membrane was incubated overnight at 4°C with primary antibodies specific to TLR2 (#bs-1019R, Bioss, 1:1000), MyD88 (#bs-1047R, Bioss, 1:1000), p-NF-κB-p65 (#bs-7343R, Bioss, 1:1000), NLRP3 (#bs-41293R, Bioss, 1:1000), caspase-1 (#bsm-52441R, Bioss, 1:1000), IL-1β (#bs-0812R, Bioss, 1:1000), and GSDMD (#bs-14287R, Bioss, 1:1000), followed by incubation with an IgG secondary antibody (#bs-10900R, Bioss, 1:5000) for 1 hour in the dark.

Techniques: Activation Assay, Western Blot, Expressing, Control

Journal: Journal of Inflammation Research

Article Title: Electroacupuncture Inhibits the Early Neuroinflammatory Cascade Triggered by TLR2 in the Prodromal Period of PD

doi: 10.2147/JIR.S585729

Figure Lengend Snippet: EA inhibits microglial pyroptosis in the pPD. ( A ) qRT-PCR was used to detect NLRP3 mRNA in the substantia nigra. ( B ) qRT-PCR was used to detect GSDMD mRNA in the substantia nigra. ( C and D ) Co-localization of Iba-1 and GSDMD in the substantia nigra was detected by immunofluorescence co-localization (magnification ×200, scale bar 50 μm). (n=3; compared with the control group, *** P <0.001; compared with the model group, ### P <0.001, ## P <0.01).

Article Snippet: The membrane was incubated overnight at 4°C with primary antibodies specific to TLR2 (#bs-1019R, Bioss, 1:1000), MyD88 (#bs-1047R, Bioss, 1:1000), p-NF-κB-p65 (#bs-7343R, Bioss, 1:1000), NLRP3 (#bs-41293R, Bioss, 1:1000), caspase-1 (#bsm-52441R, Bioss, 1:1000), IL-1β (#bs-0812R, Bioss, 1:1000), and GSDMD (#bs-14287R, Bioss, 1:1000), followed by incubation with an IgG secondary antibody (#bs-10900R, Bioss, 1:5000) for 1 hour in the dark.

Techniques: Quantitative RT-PCR, Immunofluorescence, Control

Journal: Journal of Inflammation Research

Article Title: Electroacupuncture Inhibits the Early Neuroinflammatory Cascade Triggered by TLR2 in the Prodromal Period of PD

doi: 10.2147/JIR.S585729

Figure Lengend Snippet: EA intervention affects the expression of TLR2 in different regions of pPD, inhibits microglial activation, regulates the TLR2/NF-κB/NLRP3 pathway, and suppresses pyroptosis of microglia.

Article Snippet: The membrane was incubated overnight at 4°C with primary antibodies specific to TLR2 (#bs-1019R, Bioss, 1:1000), MyD88 (#bs-1047R, Bioss, 1:1000), p-NF-κB-p65 (#bs-7343R, Bioss, 1:1000), NLRP3 (#bs-41293R, Bioss, 1:1000), caspase-1 (#bsm-52441R, Bioss, 1:1000), IL-1β (#bs-0812R, Bioss, 1:1000), and GSDMD (#bs-14287R, Bioss, 1:1000), followed by incubation with an IgG secondary antibody (#bs-10900R, Bioss, 1:5000) for 1 hour in the dark.

Techniques: Expressing, Activation Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) level (pg/ml) in patients with HCV-related liver disease and healthy controls [Data are expressed as median (interquartile range), 1040 (395) pg/ml vs 695 (183) pg/ml respectively, χ 2 = 23.888, P < 0.001, the Mood’s median test].

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Immunohistochemical staining of liver tissues from patients with hepatitis C virus-related liver disease. ( A ) METAVIR stage F1 with mild activity (A1), and no steatosis. Moderate brown positive NLRP3 (nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3) staining of hepatocytes is seen (staining score 4, low expression) (anti-NLRP3 × 200); ( B ) High power view demonstrating brown cytoplasmic NLRP3 expression in hepatocytes (anti-NLRP3 × 400); ( C ) METAVIR stage F4 (cirrhosis) with moderate activity (A2), and steatosis. Strong NLRP3 expression is seen (staining score 12, high expression) (anti-NLRP3 × 200); ( D ) METAVIR stage F2, with moderate activity (A2). Weak brown positive Caspase-1 (CASP1) staining of hepatocytes is observed (staining score 1) (anti-CASP1 × 200). ( E ) METAVIR stage F3 with moderate activity (A2), and no steatosis. Moderate brown positive CASP1 staining of hepatocytes is seen (staining score 2) (anti-CASP1 × 200); and ( F ) METAVIR stage F4 (cirrhosis). Strong brown positive CASP1 staining of hepatocytes (staining score 3) (anti-CASP1 × 200).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Immunohistochemical staining, Staining, Virus, Activity Assay, Binding Assay, Expressing

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Statistical correlations between ( A ) hepatic nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) and Caspase-1 (CASP1) staining scores (r = 0.810, P < 0.001), ( B ) serum NLRP3 levels (pg/ml) and hepatic NLRP3 final staining score (r = 0.749, P < 0.001), and ( C ) serum NLRP3 levels (pg/ml) and hepatic CASP1 staining score (r = 0.577, P = 0.001) in patients with hepatitis C virus-related liver disease (n = 34).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Binding Assay, Staining, Virus

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Statistical correlations between serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) levels (pg/ml), hepatic NLRP3 expression, and other parameters in patients with hepatitis C virus (HCV)-related liver disease.

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Expressing, Virus, Staining, Activity Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Serum levels (pg/ml) and hepatic expression of nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) in: ( A & B ) patients with severe necroinflammation (A3) vs patients with mild/moderate necroinflammation (A1-A2) ( P < 0.001 and P = 0.013 respectively), ( C & D ) patients with advanced fibrosis/cirrhosis (F3-F4) vs patients with early fibrosis (F1-F2) ( P < 0.001 for both), and ( E & F ) patients with significant steatosis (grade 2–3) vs patients with non-significant steatosis (grade 0–1) ( P < 0.001 and P = 0.001 respectively).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Expressing, Binding Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: The diagnostic performance of serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) (pg/ml) in determining the severity of liver necroinflammation, fibrosis, and steatosis.

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Diagnostic Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Receiver operating characteristic curve showing the sensitivity and specificity of ( A ) serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) (pg/ml), serum aspartate aminotransferase (AST) (U/L), and serum alanine aminotransferase (ALT) (U/L) in discriminating severe liver necroinflammation (A3) from mild/moderate liver necroinflammation (A1-A2) (area under the curve (AUC) = 0.951 vs 0.676, P = 0.032 and 0.692, P = 0.034 respectively), ( B ) serum NLRP3 (pg/ml), aspartate aminotransferase to platelet ratio index (APRI), and Fibrosis-4 index (FIB-4) in discriminating advanced liver fibrosis/cirrhosis (F3-F4) from early liver fibrosis (F1-F2) (AUC = 0.971 vs 0.754, P = 0.036 and 0.806, P = 0.075 respectively), ( C ) serum NLRP3 (pg/ml) in discriminating significant steatosis (grade 2–3) from non-significant steatosis (grade 0–1) (AUC = 0.917, P < 0.001).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: Distinct SARS-CoV-2 specific NLRP3 and IL-1β responses in T cells of aging patients during acute COVID-19 infection

doi: 10.3389/fimmu.2023.1231087

Figure Lengend Snippet: SPIKE-specific T cell-IL-1β expression is reduced in COVID+ patients aged ≥ 61. PBMCs from age ≤ 60 (n=20) and aged ≥ 61 (n=16) COVID+ patients were stimulated with SPIKE peptides in vitro for 12 h to assess COVID-specific responses in T cells. Representative contour plots showing % of PD-1 and IFN-γ expressing (A, B) , T-bet (D) , NLRP3 (E) , of CD4+ (above) and CD8+ (below) cells. Statistics showing % IFN-γ/ %PD-1 ratios (C) and NLRP3 expression (F) , in CD4+ (top) and CD8+ cells (bottom) in PBMCs from subjects aged ≤ 60 and ≥ 61 in vitro. Median values ± SEM are plotted. (Mann-Whitney U test; Two-tailed). ***<0.0005, ****<0.00005.

Article Snippet: NLRP3-PE (bs-10021R-PE) is from Bioss (MN, US).

Techniques: Expressing, In Vitro, MANN-WHITNEY, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Distinct SARS-CoV-2 specific NLRP3 and IL-1β responses in T cells of aging patients during acute COVID-19 infection

doi: 10.3389/fimmu.2023.1231087

Figure Lengend Snippet: IL-1β expression positively correlates with IFN-γ and NLRP3 expression in SPIKE-activated T cells in COVID+ patients. Spearman Correlation (r) analysis showing positive correlation between IL-1β and IFN-γ in CD4 + T cells (A) and CD8 + T cells (B) , and NLRP3 and IFN-γ in CD4 + T cells (C) , and CD8 + T cells (D) . Negative correlation between cytokines and age in CD4 + T cells (E, G) and CD8 + T cells (F, H) . *P<0.05, **<0.005, ***<0.0005, ****<0.00005.

Article Snippet: NLRP3-PE (bs-10021R-PE) is from Bioss (MN, US).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Distinct SARS-CoV-2 specific NLRP3 and IL-1β responses in T cells of aging patients during acute COVID-19 infection

doi: 10.3389/fimmu.2023.1231087

Figure Lengend Snippet: Cytokine dysregulation is not observed in aged monocytes (aged ≥ 61) in SPIKE-activated PBMCs from COVID-infected individuals. PBMCs from COVID+ patients aged ≤ 60 and ≥ 61 were stimulated with SPIKE for 12 h as in Figure 2 . Representative contour plots (A) showing % of IL-1β (x-axis) and IFN-γ (y-axis) gating on CD14 + monocytes and statistics of % IL-1β + cells in COVID+ patients aged ≤ 60 and ≥ 61 (B) . Histogram plots showing NLRP3 expression (C) and statistical analysis of % of NLRP3 + cells in COVID+ patients (D) . Median values ± SEM are plotted. (Mann-Whitney U test; Two-tailed). NS, non significant.

Article Snippet: NLRP3-PE (bs-10021R-PE) is from Bioss (MN, US).

Techniques: Infection, Expressing, MANN-WHITNEY, Two Tailed Test

Journal: Microorganisms

Article Title: Transcriptomic and Immunopathological Profiles of Inflammasomes in Different Clinical Forms of American Cutaneous Leishmaniasis

doi: 10.3390/microorganisms13050980

Figure Lengend Snippet: Boxplots showing normalized gene expression data from RNA-seq of lesions in patients with different clinical forms of American Cutaneous Leishmaniasis caused by L. (L.) amazonensis and L. (V.) braziliensis. ADCL: Anergic Diffuse Cutaneous Leishmaniasis, caused by L. (L.) amazonensis ; LCL (La): Localized Cutaneous Leishmaniasis, caused by L. (L.) amazonensis ; LCL (Lb): Localized Cutaneous Leishmaniasis, caused by L. (V.) braziliensis ; and MCL: Mucocutaneous Leishmaniasis caused by L. (V.) braziliensis. A: NLRP3; B: AIM2; C: NLRP12; D: NLRC4; E: CASP1; F: CASP5; G: GSDMD; H: IL-18; I: IL-1 β. * = p < 0.05; ** = p < 0.001 and *** = p < 0.0001.

Article Snippet: The sections were incubated with primary antibodies anti-NLRP3 (polyclonal, E-AB-70161, Elabscience, Houston, TX, USA, 1:600), anti-AIM-2 (polyclonal, E-AB-10974, Elabscience, 1:100), anti-NLRC4 (polyclonal, E-AB-67860, Elabscience, 1:100), anti-NLRP12 (polyclonal, E-AB-66298, Elabscince, 1:500), anti-caspase-1 (monoclonal, G6231-3D2, Sigma Aldrich, St. Louis, MO, USA, 1:500), anti-caspase-5 (monoclonal, MAA770Hu22, Cloud-clone Corp., Katy, TX, USA, 1:100), anti-GSDMD (monoclonal, E-AB-81473, Elabscience, 1:200), anti-IL-1β (polyclonal, ab2105, Abcam, Cambridge, MA, USA, 1:800), and anti-IL-18 (polyclonal, ab68435, Abcam, 1:2000).

Techniques: Gene Expression, RNA Sequencing

Journal: Microorganisms

Article Title: Transcriptomic and Immunopathological Profiles of Inflammasomes in Different Clinical Forms of American Cutaneous Leishmaniasis

doi: 10.3390/microorganisms13050980

Figure Lengend Snippet: Immunohistochemistry reactions on histological sections of biopsies from patients with different clinical forms of ACL using anti-NLRP3, anti-AIM-2, anti-NLRP12, anti-NLRC4, anti-caspase-1, anti-caspase-5, anti-GSDMD, anti-IL-1β, and anti-IL-18 antibodies. ADCL: Anergic Diffuse Cutaneous Leishmaniasis caused by L. (L.) amazonensis ; LCL (La): Localized Cutaneous Leishmaniasis caused by L. (L.) amazonensis ; LCL (Lb): Localized Cutaneous Leishmaniasis caused by L. (V.) braziliensis ; MCL: Mucocutaneous Leishmaniasis caused by L. (V.) braziliensis . Red arrows indicate immunostained cells.

Article Snippet: The sections were incubated with primary antibodies anti-NLRP3 (polyclonal, E-AB-70161, Elabscience, Houston, TX, USA, 1:600), anti-AIM-2 (polyclonal, E-AB-10974, Elabscience, 1:100), anti-NLRC4 (polyclonal, E-AB-67860, Elabscience, 1:100), anti-NLRP12 (polyclonal, E-AB-66298, Elabscince, 1:500), anti-caspase-1 (monoclonal, G6231-3D2, Sigma Aldrich, St. Louis, MO, USA, 1:500), anti-caspase-5 (monoclonal, MAA770Hu22, Cloud-clone Corp., Katy, TX, USA, 1:100), anti-GSDMD (monoclonal, E-AB-81473, Elabscience, 1:200), anti-IL-1β (polyclonal, ab2105, Abcam, Cambridge, MA, USA, 1:800), and anti-IL-18 (polyclonal, ab68435, Abcam, 1:2000).

Techniques: Immunohistochemistry

Journal: Heliyon

Article Title: Exposing kinetic disparities between inflammasome readouts using time-resolved analysis

doi: 10.1016/j.heliyon.2024.e32023

Figure Lengend Snippet: Temporal association between speck formation, cytokine release and LDH leakage varies with activating signal. After triggering NLRP3 inflammasome activation with either ATP (A), MSU (B) or nigericin (C) in PMA-differentiated, LPS-primed THP-1 cells, the temporal association of ASC-speck count, extracellular IL-1β concentration, extracellular IL-18 concentration, and extracellular LDH was assessed. ASC-GFP specks were imaged by fluorescent microscopy and automatically quantified using the Weka segmentation plugin for ImageJ. IL-1β concentration and IL-18 concentration were quantified using the MSD® U-PLEX Platform. LDH was quantified using the CyQuant™ LDH Cytotoxicity Assay Kit. Speck count, IL-1β concentration and IL-18 concentration are shown as mean (solid line, dark dashed line and light dashed line, respectively) ± SEM (shaded area). LDH is shown as individual data points (red dots) with the mean (black dash). Created with BioRender.com. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Cells were washed twice with PBS for 5 min and permeabilized with 0.1 % Triton X-100, 1 % FBS in PBS for 10 min, followed by blocking with 5 % FBS in PBST for 1 h. The blocking solution was replaced with rabbit polyclonal anti-NLRP3 antibodies (Merck) diluted 1:1000 in 1 % BSA in PBST and incubated at 4 °C overnight.

Techniques: Activation Assay, Concentration Assay, Microscopy, CyQUANT Assay, LDH Cytotoxicity Assay