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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: Reduced glutathione as a physiological co-activator in the activation of peptidylarginine deiminase

Fig. 4

Release of catalytically active peptidylarginine deiminase (PAD) from PMA-stimulated granulocytes. a Microtitre plates coated with human fibrinogen were incubated for 3 h at RT with blood leucocytes resuspended in RPMI medium containing 5 % normal human serum with or without 25 mM EDTA. Cells were stimulated with PMA (15 nM) and subsequent citrullination of fibrinogen was measured using mAb 20B2 as the detecting antibody. Average and upper range of duplicate optical density (OD) measurements shown for three donors. b Enzymatic activity of PMA-stimulated leucocytes from 10 individual donors was examined in a similar manner in the presence and absence of exogenous rhPAD2 (300 ng/ml) and/or 1 mM dithiothreitol (DTT). Shown are averages of duplicate measurements after subtraction of background values obtained in the presence of EDTA. c Using the same procedure, mononuclear cells (MNCs) from donor 1 in a were assessed. Average and upper range of duplicate OD measurements are shown. PMA phorbol 12-myristate 13-acetate

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