10 ng ml−1 of lps  (PeproTech)

Journal: Nature

Article Title: Clonal hematopoiesis and risk of chronic liver disease

doi: 10.1038/s41586-023-05857-4

Figure Lengend Snippet: a, Association of the IL6R germline mutation resulting in p.Asp358Ala with chronic liver disease in individuals with CHIP (variant allele fraction ≥ 10%) versus individuals without CHIP. b, Unsupervised hierarchical clustering of differentially regulated genes in sorted liver macrophages from B6.SJL mice transplanted with Tet2−/− (n = 4) or control vavCre+ (WT; n = 4) bone marrow cells and fed CDAHFD for 11 weeks. c, After 19 weeks of CDAHFD, Tet2−/−-transplanted (n = 12) or control vavCre+-transplanted (WT; n = 14) mice were bled and serum was obtained for cytokine measurements. Statistical analysis was performed using two-tailed unpaired t-test (IL-6, CXCL1) or Mann–Whitney test (CCL22, CCL17) with Bonferroni correction for multiple hypothesis testing. d, Bone marrow-derived macrophages from Tet2−/− (n = 6), Tet2−/−Nlrp3−/− (n = 4) or control vavCre+ (WT, n = 7) mice were primed with low-dose lipopolysaccharide (LPS) for 2 h and stimulated with palmitic acid or cholesterol monohydrate crystals as indicated for 6 h. Statistical analysis was performed using two-way ANOVA. For c and d, individual measurements, mean and standard deviation from two independent experiments are shown.

Article Snippet: Macrophages were exposed to 10 ng ml −1 LPS (Sigma) for 2 h and stimulated for 6 h with palmitic acid (300 μM; Sigma) or cholesterol monohydrate crystals (200 μg ml −1 ; Sigma) prepared as previously described 38 .

Techniques: Mutagenesis, Variant Assay, Two Tailed Test, MANN-WHITNEY, Derivative Assay, Standard Deviation

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Endotoxin tolerance and low activation of TLR-4/NF-κB axis in monocytes of COVID-19 patients

doi: 10.1007/s00109-023-02283-x

Figure Lengend Snippet: Analysis of systemic levels of LPS ( A ), soluble CD14 ( B ), cytokines concentrations ( C , IL-1β; E , IL-6; F , IL-10; G , TNF-α; H , MCP-1), and prostaglandin E2 ( D ) in the plasma of healthy controls, mild COVID-19, and severe COVID-19 patients. Representative dot plot of CD14 + HLA-DR low/+ ( I ), the expression of TLR-4 and NF-κB p65 in CD14 + HLA-DR + ( J ), and the expression of TLR-4 and NF-κB p65 in CD14 + HLA-DR low ( K ) monocytes are showed. Finally, whole blood was used to evaluate the peripheral frequency of CD14 + HLA-DR + ( L ) and CD14 + HLA-DR low ( M ), the expression of TLR-4 ( N ) and NF-κB p65 ( O ) in CD14 + HLA-DR + , and the expression of TLR-4 ( P ) and NF-κB p65 ( Q ) in CD14 + HLA-DR. low in healthy controls, mild COVID-19, and severe COVID-19 patients. Between-group comparison performed through one-way ANOVA followed by Bonferroni’s post hoc. ( p < 0.05). * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Whole blood samples of controls, mild COVID-19, and severe COVID-19 patients were aliquoted in tubes to a volume of 1 mL and incubated (5%CO 2 , 37 °C) with two different LPS concentrations (1 and 10 ng/mL, Sigma-Aldrich, USA) or without immunogenic stimulus for 2 h. The phenotype of monocytes was evaluated by flow cytometry.

Techniques: Expressing

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Endotoxin tolerance and low activation of TLR-4/NF-κB axis in monocytes of COVID-19 patients

doi: 10.1007/s00109-023-02283-x

Figure Lengend Snippet: The production of IL-1β ( A ), IL-6 ( B ), IL-10 ( C ), PGE-2 ( D ), MCP-1 ( E ), and TNF-α ( F ) by ex vivo LPS-stimulated whole blood of healthy controls and mild and severe COVID-19 patients. Whole blood samples of controls, mild COVID-19, and severe COVID-19 patients were aliquoted in tubes to a volume of 1 mL and incubated (5%CO 2 , 37 °C) with two different LPS concentrations (1 and 10 ng/mL, Sigma-Aldrich, USA) or without immunogenic stimulus for 2 h. The supernatant concentrations of interleukin (IL)-6, IL-10, C–C motif ligand 2 (CCL2), and tumor necrosis factor-alpha (TNF-α) were analyzed by multiplex assays. The supernatant Prostaglandin E2 and IL-1β concentrations were analyzed by enzyme‐linked immunosorbent assay (ELISA). The production of cytokines was evaluated by the % of difference compared to the non-stimulated condition. **( p < 0.01) and ***( p < 0.001) denote statistical difference ( p < 0.01)

Article Snippet: Whole blood samples of controls, mild COVID-19, and severe COVID-19 patients were aliquoted in tubes to a volume of 1 mL and incubated (5%CO 2 , 37 °C) with two different LPS concentrations (1 and 10 ng/mL, Sigma-Aldrich, USA) or without immunogenic stimulus for 2 h. The phenotype of monocytes was evaluated by flow cytometry.

Techniques: Ex Vivo, Incubation, Multiplex Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Endotoxin tolerance and low activation of TLR-4/NF-κB axis in monocytes of COVID-19 patients

doi: 10.1007/s00109-023-02283-x

Figure Lengend Snippet: The frequencies of CD14 + HLA-DR. low ( A ) monocytes expressing TLR-4 ( B ) and NF-κB ( C ) and CD14 + HLA-DR + ( D ) expressing TLR-4 ( E ) and NF-κB ( F ) in ex vivo LPS-stimulated whole blood of healthy controls and mild and severe COVID-19 patients. Whole blood samples of controls, mild COVID-19, and severe COVID-19 patients were aliquoted in tubes to a volume of 1 mL and incubated (5%CO 2 , 37 °C) with two different LPS concentrations (1 and 10 ng/mL, Sigma-Aldrich, USA) or without immunogenic stimulus for 2 h. The phenotype of monocytes was evaluated by flow cytometry. The frequencies of each cell subtype were evaluated by the % of difference compared to the non-stimulated condition. *( p < 0.05), **( p < 0.01), and ***( p < 0.001) denote statistical difference ( p < 0.01)

Article Snippet: Whole blood samples of controls, mild COVID-19, and severe COVID-19 patients were aliquoted in tubes to a volume of 1 mL and incubated (5%CO 2 , 37 °C) with two different LPS concentrations (1 and 10 ng/mL, Sigma-Aldrich, USA) or without immunogenic stimulus for 2 h. The phenotype of monocytes was evaluated by flow cytometry.

Techniques: Expressing, Ex Vivo, Incubation, Flow Cytometry