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Figure 4: Hypercitrullination in other leukocytes. (a) Neutrophils, (b) PBMC, (c) monocytes, and (d) T cells were isolated from the same donor and cultured with or without 1 μM ionomycin, 100 nM PMA, and 0.005% Triton-X for one hour in medium with 2 mM CaCl2. Citrullination was detected with the F95 antibody (upper panel) and equal loading verified with anti-β-actin (lower panel). The shown blots are representative of four different donors. (e) B cells were isolated and cultured with or without 1 μM ionomycin in 2 mM CaCl2 medium. The indicated number of cell equivalents was loaded and citrullinated proteins were detected by F95 antibody (left panel) or anti-modified citrulline clone antibody (AMC) (right panel). The shown blots are representative of five different donors. (f) NK cells, myeloid dendritic cells (mDC), and plasmacytoid dendritic cells (pDC) were incubated in medium with 2 mM CaCl2 medium alone or with 1 μM ionomycin for 1 h, as indicated. Citrullination was detected with the F95 antibody (upper panel) and equal loading verified with anti-β-actin (lower panel). The blot is representative of three different donors.