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. 2020 Apr 17;11:573. doi: 10.3389/fmicb.2020.00573

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Copyright © 2020 Kamoshida, Akaji, Takemoto, Suzuki, Sato, Kai, Hibino, Yamaguchi, Kikuchi-Ueda, Nishida, Unno, Tansho-Nagakawa, Ubagai, Miyoshi-Akiyama, Oda and Ono.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Measurements of inflammatory response molecules secreted by neutrophils. ROS and superoxide expression levels in neutrophils challenged by bacteria (ATCC 19606: WT, colistin-resistant A. baumannii: CRAb-5) were measured by flow cytometry and a total ROS/superoxide detection kit (A). PMA (100 nM)-stimulated neutrophils served as positive controls. Data are means ± SD; n = 5 per group. ^∗∗∗P < 0.001. (B) Inflammatory cytokines such as interleukin (IL)-8, tumor necrosis factor (TNF), IL-1β, and IL-6 produced by neutrophils in response to bacterial stimulation were measured by the cytometric bead array system. Supernatants were obtained by co-culturing neutrophils and bacteria for 4 h. The supernatants of neutrophils alone stimulated with PMA (100 nM) served as positive controls. Data are means ± SD; n = 4 per group. ^∗∗∗P < 0.001.