Journal: Cells

Article Title: Aggregated Tau-PHF6 (VQIVYK) Potentiates NLRP3 Inflammasome Expression and Autophagy in Human Microglial Cells

doi: 10.3390/cells10071652

Figure Lengend Snippet: List of antibodies used for immunofluorescence staining.

Article Snippet: NLRP3 , Bioss, Woburn, MA, USA , bs-10021R , rabbit , 1:500.

Techniques: Immunofluorescence, Staining

Journal: Journal of Immunology Research

Article Title: TLR4-NLRP3-GSDMD-Mediated Pyroptosis Plays an Important Role in Aggravated Liver Injury of CD38 −/− Sepsis Mice

doi: 10.1155/2021/6687555

Figure Lengend Snippet: Escherichia coli can induce liver injury in mice. WT mice were injected with PBS or 3 × 10 8 cfu/ml E. coli intraperitoneally and sacrificed 3 hours later. Liver pathological injuries were observed with hematoxylin and eosin staining (a–d). The yellow arrows indicate edema, the blue arrows indicate inflammatory cell infiltration, the black arrows indicate punctate necrosis, and the green arrows indicate binucleate hepatocytes. Bacteria from the liver of PBS- or E. coli -stimulated WT mice were cultured in MH medium (e, f) overnight and identified by Gram staining (g). The serum AST (h) and ALT (i) concentrations were determined by using the detection kits. The mRNA of liver inflammatory cytokines TLR4 (j), NLRP3 (k), IL-1 β (l), and IL-18 (m) of PBS- or 3 × 10 8 cfu/ml E. coli -stimulated mice were measured by RT-qPCR. Data are presented as means ± standard deviation. Statistical significance was determined by the paired t -test ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01).

Article Snippet: The primary antibodies anti-TLR4 rabbit mAb (1 : 500) (CST, USA), anti-TRIF rabbit mAb (1 : 1000) (Proteintech, USA), anti-MyD88 mouse mAb (1 : 2000) (Proteintech, USA), anti-NF- κ B p65 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-NF- κ B p65 rabbit mAb (1 : 500) (CST, USA), anti-IL-6 mouse mAb (1 : 2000) (Proteintech, USA), anti-iNOS rabbit mAb (1 : 1000) (CST, USA), anti-BAX rabbit mAb (1 : 5000) (Proteintech, USA), anti-NLRP3 rabbit mAb (1 : 500) (Boster, China), anti-ASC rabbit mAb (1 : 500) (Affinity, China), anti-caspase-1 rabbit mAb (1 : 500) (Abcam, UK), anti-IL-1 β rabbit mAb (1 : 1000) (CST, USA), anti-IL-18 rabbit mAb (1 : 1000) (Affinity, China), anti-caspase-3 rabbit mAb (1 : 500) (CST, USA), anti-GSDMD rabbit mAb (1 : 500) (Affinity, China), anti-ERK1/2 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-ERK1/2 rabbit mAb (1 : 2000) (CST, USA), and anti-GAPDH rabbit mAb (1 : 5000) (Proteintech, USA) were used.

Techniques: Injection, Staining, Bacteria, Cell Culture, Quantitative RT-PCR, Standard Deviation

Journal: Journal of Immunology Research

Article Title: TLR4-NLRP3-GSDMD-Mediated Pyroptosis Plays an Important Role in Aggravated Liver Injury of CD38 −/− Sepsis Mice

doi: 10.1155/2021/6687555

Figure Lengend Snippet: The expression levels of pyroptosis-related markers were detected by Western blot. The expressions of liver pyroptosis proteins in WT, CD38 −/− , and CD38 −/− TLR4 mut mice were detected at 3 hours after E. coli stimulation by Western blot. The expressions of NLRP3, ASC, procaspase-1, cleaved caspase-1, IL-1 β , IL-18, procaspase-3, and cleaved caspase-3 were measured (a). And relative levels of NLRP3 to GAPDH (b), ASC to GAPDH (c), cleaved to procaspase-1 (d), IL-1 β to GAPDH (e), IL-18 to GAPDH (f), and cleaved to procaspase-3 (g) were analyzed by ImageJ software. Data are presented as means ± standard deviation. Statistical significance was determined by one-way ANOVA ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01).

Article Snippet: The primary antibodies anti-TLR4 rabbit mAb (1 : 500) (CST, USA), anti-TRIF rabbit mAb (1 : 1000) (Proteintech, USA), anti-MyD88 mouse mAb (1 : 2000) (Proteintech, USA), anti-NF- κ B p65 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-NF- κ B p65 rabbit mAb (1 : 500) (CST, USA), anti-IL-6 mouse mAb (1 : 2000) (Proteintech, USA), anti-iNOS rabbit mAb (1 : 1000) (CST, USA), anti-BAX rabbit mAb (1 : 5000) (Proteintech, USA), anti-NLRP3 rabbit mAb (1 : 500) (Boster, China), anti-ASC rabbit mAb (1 : 500) (Affinity, China), anti-caspase-1 rabbit mAb (1 : 500) (Abcam, UK), anti-IL-1 β rabbit mAb (1 : 1000) (CST, USA), anti-IL-18 rabbit mAb (1 : 1000) (Affinity, China), anti-caspase-3 rabbit mAb (1 : 500) (CST, USA), anti-GSDMD rabbit mAb (1 : 500) (Affinity, China), anti-ERK1/2 rabbit mAb (1 : 1000) (CST, USA), anti-phospho-ERK1/2 rabbit mAb (1 : 2000) (CST, USA), and anti-GAPDH rabbit mAb (1 : 5000) (Proteintech, USA) were used.

Techniques: Expressing, Western Blot, Software, Standard Deviation

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