Colchicine inhibits the MSU-induced internalization of MICL in human neutrophils. Freshly isolated human neutrophils were treated with colchicine (10 µM) (+) or dimethyl sulfoxide (DMSO) (-) for 30 min at 37°C and then incubated with (A) MSU (1 mg/ml) for 20 min at 37°C or (B) 50C1 for 5 min at 37°C. The stimulations were terminated by transferring the tubes to an ice bath, followed by centrifugation at 400 × g for 2 min at 4°C. The cell pellets were washed in cold HBSS containing 0.005% BSA and incubated with (A) 50C1 for 30 min, followed by incubation with FITC-labeled goat antimouse Fcγ-specific IgG (diluted 1:100 in HBSS/BSA) for 30 min on ice, or (B) FITC-labeled goat anti-mouse Fcγ-specific IgG (diluted 1:100 in HBSS/BSA) for 30 min on ice. Cells were then washed twice in HBSS/BSA and analyzed by flow cytometry using the FACSCanto II flow cytometer (BD Biosciences). This graph is a compilation of 7 independent experiments in (A) and 3 independent experiments in (B).
Gagné et al. Arthritis Research & Therapy 2013 15:R73 doi:10.1186/ar4250