Journal: iScience

Article Title: Annexin A1 exerts analgesic effect in a mouse model of medication overuse headache

doi: 10.1016/j.isci.2023.108153

Figure Lengend Snippet:

Article Snippet: The stock solutions of the formyl peptide receptor (FPR) antagonist Boc-MLF TFA (MedChemExpress LLC, HY-103473A, Shanghai, China) were prepared by dissolving it in dimethyl sulfoxide.

Techniques: Recombinant, SYBR Green Assay, Lysis, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Western Blot, Software, Microscopy, Real-time Polymerase Chain Reaction, Imaging, Injection

Journal: Scientific Reports

Article Title: Formylpeptide receptors are critical for rapid neutrophil mobilization in host defense against Listeria monocytogenes

doi: 10.1038/srep00786

Figure Lengend Snippet: (a) Chemotactic activity of Listeria -derived peptide fMIVIL for mouse neutrophils. * significantly increased cell migration in response to the peptide as compared with medium control (0) ( p = 0.0007). (b) Chemotactic activity of Listeria lysate for HEK293 cells transfected with Fprs. * significantly increased cell chemotaxis in response to Listeria lysate as compared with medium control (0) ( p = 0.004). (c) Migration of mouse neutrophils to Listeria lysate. * significantly increased chemotaxis response of neutrophils to lysates as compared with medium control (0) ( p = 0.003). (d) Listeria lysate-induced phosphorylation of Erk1/2 in WT mouse neutrophils in the presence or absence of Fpr antagonists and a TLR2 neutralizing antibody. (e) Semiquantitative analysis of phosphorylated Erk1/2. * significantly decreased Erk1/2 phosphorylation compared with cells stimulated with Listeria lysate at 1: 10 dilution in the absence of Fpr antagonists or TLR2 antibody. Results are from 1 experiment out of 3 performed. (f) Induction of Erk1/2 phosphorylation in WT and Fpr1/2 −/− mouse neutrophils by Listeria lysate (at 1:10 dilution) in the presence or absence of an Fpr antagonist Boc-2. (g) Semiquantitative analysis of phosphorylated Erk1/2. # significantly reduced Erk1/2 phosphorylation in Fpr1/2 −/− mouse neutrophils as compared with WT mouse neutrophils. * significantly decreased Erk1/2 phosphorylation in WT mouse neutrophils stimulated with Listeria lysate in the presence of Fpr antagonist Boc-2 as compared with cells stimulated with Listeria lysate alone ( p = 0.001).

Article Snippet: The other reagents and sources were: Rat anti-mouse Ly6G, F4/80, goat anti-rat serum antibodies, mouse CXCL1 and CXCL2 ELISA kits (eBioscience, San Diego, CA); ,6-diamidino-2-phenylindole (DAPI, Molecular Probe, Eugene, OR); Alexa Fluora 488-labelled goat anti-rat IgG , ProLong antifade reagent, amplex red hydrogen peroxide/peroxidase assay kit, Qtracker® Cell Labeling Kits (Invitrogen, Eugene, OR); rat anti- mouse Erk1/2, rat anti-mouse phospho-Erk1/2, rat anti- mouse p38, rat anti-mouse phospho-p38, goat anti-rat HRP-IgG antibodies (Cell signaling, Beverly, MA); Boc-MLF, Boc-2, TLR2 antibody, MMK-1, CCL2, CXCL2, phorbol myristate acetate (PMA) (Tocris, Ellisville, MO); Igepal CA-630 nonionic detergent (R&D, Minneapolis, MN); Formyl-Met-Leu-Phe (fMLF), Percoll, Saponin, Casein and Thioglycollate, (Sigma-Aldrich, St. Louis, MO); Brain Heart Infusion Broth and Brain Heart Infusion Agar (BD, Franklin Lakes, New Jersey); FITC-labeled anti- Listeria polyclonal antibody (Antibodies-online, Atlanta, GA).

Techniques: Activity Assay, Derivative Assay, Migration, Transfection, Chemotaxis Assay