12-mer peptides on a 150×100 cellulose membrane in a 10×30 array  (INTAVIS Inc)

Journal: iScience

Article Title: Nouveau benzo-mimetics of 17R-Resolvin D2 are potent resolution agonists for inflammation

doi: 10.1016/j.isci.2025.114201

Figure Lengend Snippet: Benzo-17R-RvD2 mimetic in peritonitis limits PMN tissue infiltration and regulates cytokines Mice were administered with vehicle or benzo-17R-RvD2 (1 ng/mouse) together with zymosan A (1 mg/mouse, i.p.) and exudates collected at 4 h. Total leukocytes were enumerated and composition determined by flow cytometry (see B for gating strategy). (A) Exudate PMN numbers; mean ± SEM, n = 4 mice each group, ∗∗ p < 0.01. (B) Proteome analysis was carried out with inflammatory exudates from four individual mice in each group using Proteome Profiler cytokine arrays, which contains 40 cytokines, chemokines, and acute phase proteins. Orthogonal Partial Least Squares Discriminant Analysis (PLS-DA): the score plot (each dot represents the cytokine profile from each mouse) shows clustering among groups (i.e., peritonitis vs. peritonitis plus benzo-17R-RvD2). The loading plot demonstrates correlations in which the measured cytokines contribute to the cluster separation in the score plot. (C) Relative intensities of cytokines, chemokines, and acute phase proteins. Proteome arrays consist of 40 proteins. Among them 18 were present in the peritoneal exudates. Results are mean ± SEM; relative abundance from n = 4 individual mouse exudates, ∗ p < 0.05, ∗∗ p < 0.01; peritonitis plus benzo-17R-RvD2 versus peritonitis alone; Student’s t test.

Article Snippet: The supernatant (750 μl) was incubated with the pre-coated Proteome Profiler array membranes (ARY006, R&D Systems) and processed according to the manufacturer’s instructions.

Techniques: Flow Cytometry