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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 improves bacterial killing of C. rodentium independent from the NLRP3 inflammasome. (A) Raw264.7, J77^ASC−/−, or J77^scramble, cells were first treated with YVAD as indicated for a total of 3 h. Inoculation of cell culture 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 (25 mg/mL) was added to the culture media. Gentamicin (25 ug/mL) was used to eliminate non-invasive bacteria in (B) Raw264.7 cells or (C) J77^ASC−/−, and J77^scramble, and (D) cells treated with or without MCC950 (1 μM). Cell lysates were collected and grown on LB agar overnight and colonies were manually counted and expressed as CFU/mL. Values represent mean ± SEM between 3 independent experiments; *p < 0.05. YVAD (Ac-Tyr-Val-Ala-Asp-Chloromethylketone); MOI (multiplicity of infection); CFU (colony forming units).