Figure 4. AG induces oxidative shift in EMSH of Mtb in vitro and during infection.
(A) Wt Mtb-expressing Mrx1-roGFP2 was treated with lethal (10X MIC) and sub-lethal (0.2 X MIC) concentrations of AG and ratiometric sensor response was measured at indicated time points by flow cytometry. (B) PMA-differentiated THP-1 cells were infected with Mtb expressing Mrx1-roGFP2 (moi: 10) and treated with indicated concentrations of Amox in the presence of 8 µg/ml of Clav as described in Materials and methods. At the indicated time points, ~30,000 infected macrophages were analyzed by flow cytometry to quantify changes in Mtb subpopulations displaying variable EMSH as described in Materials and methods. (C) In parallel experiments, infected macrophages were lysed and bacillary load was measured by plating for CFU. Error bars represent standard deviations from the mean. *p≤0.05, **p≤0.01 and ***p≤0.001. Data are representative of at least two independent experiments done in duplicate.