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. 2018 Mar 8;8:4168. doi: 10.1038/s41598-018-22343-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Increased sensitivity of enhanced acid growth mutants to antibiotics. Wild type Mtb (WT), an eag4 mutant, and WT Mtb expressing either the wild type allele or S211R-encoding mutant allele of MT3221 (WT + pVV-MT3221-WT and WT + pVV-MT3221-S211R, respectively) were acclimated to minimal medium containing glycerol as a single carbon source for 3 days before adding either 0.6 µM rifampin (A), 20 µM isoniazid (B), or 10 µM PA-824 (C). The eag4 mutant exhibited increased sensitivity to both rifampin and isoniazid at pH 5.7 but not pH 7.0, and the strain overexpressing the S211R-encoding MT3221 mutant allele also had increased sensitivity to isoniazid at pH 5.7. Both the mutant strain and the MT3221 overexpression strains demonstrated increased sensitivity to PA-824 at pH 7.0, but no significant tolerance was observed at pH 5.7. *P < 0.05 based on a student’s t-test.