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. 2019 Oct 1;10:2296. doi: 10.3389/fimmu.2019.02296

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Copyright © 2019 Armstrong, Bording-Jorgensen, Chan and Wine.

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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Nigericin results in bacterial killing in Raw 264.7 cells through a mechanism unrelated to the NRLP3 inflammasome. Raw 264.7 cells were first treated with YVAD as indicated for a total of 3 h. Inoculation of Raw 264.7 with C. rodentium (MOI 1:10) occurred 1 h after initial addition of YVAD. 1.5 h after C. rodentium inoculation, Nigericin (20 μM) or ATP control (2.5 mM) was added to the culture media. Cells were lysed with trizol and RNA extraction and cDNA processing proceeded as described in the methods. qPCR was performed to examine NLRP3, IL-1β, IL-10, Cd40lg, Ciita, IL-12, IFNγ, P38, Nlrp4e, Tab2, TNFα, and Tnfsf11. Samples were controlled against β-actin and Rer1 expression. *p < 0.05.