Recombinant murine e selectin fc

RPMI-1640, Ammonium-Chloride-Potassium (ACK) lysing buffer, EDTA 0.5M, and HEPES 1M were purchased from Lonza (Walkersville, MD). MEM nonessential amino acids (NEAA), L-glutamine, Penicillin/streptomycin, Hanks’ Balanced Salt Solution with Ca²⁺ and Mg²⁺ (HBSS⁺) or HBSS without Ca²⁺ and Mg²⁺ (HBSS⁻), and Phosphate Buffered Saline with (PBS⁺) or without Ca²⁺ and Mg²⁺ (PBS⁻) were purchased from Corning Cellgro, Mediatech Inc. (Tewksbury, MA). Fetal bovine serum (FBS) was from Atlanta Biologicals (Oakwood, GA). Leukotriene B₄ (LTB₄) was from Cayman Chemical (Ann Arbor, MI). N-Formylmethionyl-leucyl-phenylalanine (fMLF), DNase I, Histopaques-1077 and −1119, 2,2'-azino-di-(3-ethyl)di-thiazoline-sulfonic-acid (ABTS), DuoLink^® In situ-Fluorescence kit, and proteinase inhibitor cocktail (cat#P8340) were from Millipore-Sigma (St. Louis, MO). Recombinant Protein G-Sepharose^® 4B, Hoechst 33342, CountBright counting beads, and Prolong antifade mounting agent were from Invitrogen (Carlsbad, CA). Recombinant murine E-selectin-Fc, VCAM-1-Fc and ICAM-1-Fc chimera molecules, and the polyclonal goat anti-mouse CD47 (cat#AF1866) and goat anti-human JAM-A (cat#AF1077) antibodies were from R&D Systems (Minneapolis, MN). Recombinant mouse TNF-α and IFN-γ were purchased from Peprotech (Rocky Hill, NJ). A complete list of antibodies used in this work is provided as supplemental material.