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. 2016 Jun 8;7:227. doi: 10.3389/fimmu.2016.00227

Figure 3.

Figure 3

Effect of Cl-amidine on citrullination and MPO-ANCA production in vivo. Experimental protocol (A). BALB/c mice (14-week-old female) were given i.p. injection of PMA (50 ng at days 0 and 7) and oral PTU (2.5 mg/day) for 2 weeks (p.o.: Per Os). These mice were divided into two groups. The first group of mice was given daily i.p. injection of PBS (200 μl/day) (n = 13). The other group was given daily i.p. injection of Cl-amidine (0.3 mg/200 μl PBS/day) (n = 7). Mouse urine was collected during the last 24 h using metabolic cages. Blood and tissue samples were obtained at day 14. NET formation in peritoneal tissues (B). The formalin-fixed paraffin-embedded sections of peritoneal tissues were subjected to immunohistochemistry for citrullinated histone 3. Representative photos among five random fields of view (×100) were shown. NET induction was quantified by calculating the rate of citrullinated histone 3-positive cells in polymorphonuclear cells in the five random fields of view. *p < 0.05 in Mann–Whitney U-test. Serum titers of MPO-ANCA determined by ELISA (C). **p < 0.01 in Mann–Whitney U-test.