Unlike 3HC-2 dye conjugates, 3HC-3-Tre labeling is initially localized
at the septum and poles. (A) Time-lapse microscopy of Msmeg cells
treated with 100 μM DMN-Tre, 3HC-3-Tre, 3HC-2-Tre, or 3HC-2-Glc
for 30 min revealed concentration of 3HC-3-Tre at cell septa and poles.
White arrows denote septal labeling. Scale bar: 5 μm. (B) Quantification
of Msmeg fluorescence in the presence of 100 μM (i) DMN-Tre,
(ii) 3HC-3-Tre, or (iii) 3HC-2-Tre during labeling for 30 min (left,
volume-normalized intensity) and subsequent washing with growth medium
for 1 h (right, total fluorescence). The number of cells included
in each analysis (n) is provided in each panel. Shaded
error bars represent ±1 standard deviation.