Fig. 4 Labeling of Msmeg with DMN-Tre is fast and specific and depends on Ag85A function. Labeling of Msmeg with DMN-Tre is fast and specific and depends.

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Fig. 4 Labeling of Msmeg with DMN-Tre is fast and specific and depends on Ag85A function. Labeling of Msmeg with DMN-Tre is fast and specific and depends on Ag85A function. (A) Flow cytometry mean fluorescence intensity (MFI) analysis of Msmeg labeling time course. Cells were incubated with 100 μM DMN-Tre for 20, 60, 90, or 240 min. a.u., arbitrary units. (B) Quantitative microscopy analysis of mCherry Msmeg cells after incubation with 100 μM DMN-Tre for 30 min, displaying the ratio of total area of DMN fluorescence over total area of mCherry fluorescence. (C to E) Flow cytometry MFI analysis of Msmeg cells (C) incubated for 30 min with 100 μM DMN-Tre or DMN-Glc and competition of DMN-Tre labeling (D) with free trehalose using the same labeling conditions as in (C); (E) Msmeg cells preincubated with ebselen, an Ag85 inhibitor, for 3 hours and labeled with DMN-Tre as in (C). Data are means ± SEM from at least two independent experiments. Data were analyzed by Student’s t test in (B) and one-way analysis of variance (ANOVA) test in (A) and (C) to (E) (***P < 0.001 and ****P < 0.0001; ns, not significant). Mireille Kamariza et al., Sci Transl Med 2018;10:eaam6310 Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works