(a)
(b)
Figure 2: Time-course and dose-response of hypercitrullination in human neutrophils. (a) Isolated neutrophils were cultured in medium with 2 mM CaCl2 alone or in the presence of 1 μM ionomycin, 0.005% Triton-X, or 500 ng/mL perforin for the indicated times. Citrullinated proteins were detected by anti-citrulline antibody clone F95 (upper panels) and similar loading of cell lysate was verified with anti-β-actin blots of the same filters (lower panels). (b) Isolated neutrophils were cultured in medium with 2 mM CaCl2 alone for 1 h or in the presence of the indicated concentrations of ionomycin or perforin for 30 min (upper panels). Anti-β-actin blots of the same filters (lower panels). Results are from one donor representative of three different donors.