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ΔleuD ΔpanCD M. tuberculosis was labeled +/− 6-TreAz, O-AlkTMM, or O-AzAlkTMM for 16–24 hrs or ~1 generation time, fixed, then subjected to successive rounds of CuAAC, the first with Carboxyrhodamine 110 alkyne (alkCR110), and the second with MB 543 Picolyl Azide (azMB543, see Methods for details). Schematic of labeling, (A); representative images (see Fig. S2 for additional images), (B); flow cytometry analysis, (C).
