monosodium urate msu tlrl msu  (InvivoGen)

Journal: Frontiers in Immunology

Article Title: A feedback loop sustaining neutrophil extracellular trap formation involves S100 proteins, histones, TLR2 and RAGE, and is restrained by albumin

doi: 10.3389/fimmu.2026.1774475

Figure Lengend Snippet: Effect of depleting discrete protein subsets on the ability of SD-supts to induce NETs. Human neutrophils adherent to poly-L-lysine-coated coverslips were stimulated with 1 mg/mL monosodium urate (MSU) for 2.5 h. The culture supernatant was collected and depleted of the original stimulus, yielding a stimulus-depleted culture supernatant (SD-supt). These SD-supts were then pre-cleared and immunoprecipitated using pan-histone (“histones”) or pan-S100 proteins (“S100”) antibodies as described in Methods. Alternatively, SD-supts were pre-cleared, mixed with rh sRAGE, and immunoprecipitated using anti-sRAGE antibodies (“sRAGE”). The resulting supernatants from immunodepleted SD-supts were stored and later used as a NET stimulus. Human neutrophils adherent to poly-L-lysine-coated coverslips were incubated for 4 h at 37 °C in the absence of stimuli (“unstim”) or in the presence of SD-supts that had been only pre-cleared (“isotype”) or immunodepleted of S100 proteins, histones, or sRAGE-bound proteins. NET formation was then assessed by microscopy and standardized NET indices were calculated. A representative experiment is shown (right panel), along with compiled data (mean ± s.e.m.) from at least 3 independent experiments. **, p < 0.01; ***, p< 0.001 vs the positive control; using Student’s paired t test.

Article Snippet: Among neutrophil stimuli, monosodium urate (MSU) (tlrl-msu) and ultra-pure peptidoglycan (PGN) were from Invivogen (#tlrl-pgnb3); TNFα (#210-TA) and GM-CSF (#7954-GM) were from R&D Systems; and N-formyl-methionyl-phenylalanine (fMLP) was from Millipore Sigma (#F3506).

Techniques: Immunoprecipitation, Incubation, Microscopy, Positive Control

Journal: Frontiers in Pharmacology

Article Title: Impact of dipeptidyl peptidase I and neutrophil serine proteases on neutrophil functional responses

doi: 10.3389/fphar.2026.1689804

Figure Lengend Snippet: Genetic ablation of DPP1 has no impact on neutrophil migration or pro-inflammatory cytokine generation in vivo. (A) WT or DPP1-KO animals were instilled intratracheally with LPS or PBS control; after 24 h, BALF was collected, and infiltrating cells were analyzed by flow cytometry. (B) Sterile air was injected dorsally into WT or DPP1-KO animals to form a dorsal air pouch. Thereafter, MSU or PBS control was injected into the air pouch; after 5 h, the pouch was washed with PBS and infiltrating cells were analyzed by flow cytometry. Data is mean ± SEM from 6 to 10 animals in each group. Statistical analysis by Student’s t-test (A) and two-way ANOVA (B) .

Article Snippet: MSU crystals (InvivoGen, Cat# tlrl-msu) at 5 mg/mL in PBS were used to induce inflammation in the dorsal air pouch model. On day 0, mice were weighed, shaved, and injected subcutaneously with 5 mL sterile air over the dorsum to form an air pouch.

Techniques: Migration, In Vivo, Control, Flow Cytometry, Sterility, Injection