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. 2019 Aug 20;10:1922. doi: 10.3389/fmicb.2019.01922

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Copyright © 2019 Mouton, Heunis, Dippenaar, Gallant, Kleynhans and Sampson.

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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Fluorescence dilution demonstrates reduced replication of M. tuberculosisΔleuDΔpanCD under acid stress in comparison to M. tuberculosis H37Rv. M. tuberculosis H37Rv and M. tuberculosisΔleuDΔpanCD containing pTiGc was cultured in the presence of 4 mM theophylline (Theo), before removal of theophylline and exposure to pH 4.5 and pH 6.5 for 48 h prior to analyses by flow cytometry. Flow cytometry histograms demonstrate increased TurboFP635 fluorescence intensity in acidic media (pH 4.5, red), compared to normal media (pH 6.5, green) in M. tuberculosisΔleuDΔpanCD (A) after 48 h. In contrast, M. tuberculosis H37Rv demonstrated similar, reduced, TurboFP635 fluorescence intensity after 48 h in acidic media (pH 4.5, red) and control media (pH 6.5, green) (B). Representative examples of three independent biological repeats are shown.