MICL modulates the MSU-induced tyrosine phosphorylation pattern in human neutrophils. Human neutrophils (20 × 106 cells/ml) were preincubated with anti-MICL antibody (clone 50C1, 2 μg/ml) (+) or IgG2a isotype (-) for 5 min and then washed. Next, cells were stimulated with MSU (1 mg/ml) at 37°C. Aliquots of the cell suspension were stopped at the indicated times by transferring them directly into the same volume of 2× boiling modified Laemmli sample buffer. Whole-cell lysates were probed by Western blotting for tyrosine phosphorylation (clone 4G10, upper panel) and flotillin-1 (lower panel), the loading control. This result is representative of three independent experiments.
Gagné et al. Arthritis Research & Therapy 2013 15:R73 doi:10.1186/ar4250