Fig. 2.
DivIVAMsm concentration correlates with the site of growth. (A) Time-lapse microscopy of the M. smegmatis strain expressing both DivIVA–Dendra (pXM05) and endogenous native DivIVA. Representative images were recorded at the indicated time (minutes). Asterisks show new poles, and arrows show old poles. (Scale bar, 2 μm.) (B and C) M. smegmatis cells expressing DivIVA–Dendra (pXM05) and endogenous DivIVA were nonspecifically labeled on their surface with the amine-reactive dye (AF-546), and unlabeled extensions were allowed to grow for 60 min. Representative image (B) and fluorescence quantification over the long axis of 16 cells (C) are shown (see also Fig. S2C). (Scale bar, 5 μm.) (D) Time-lapse microscopy of the divIVA–egfp M. smegmatis strain that only expresses the tagged copy of DivIVA at the gene locus. Representative images were recorded at the indicated time (hours). Asterisks show new poles, and arrows show old poles. (Scale bar, 2 μm.) (E and F) divIVA–egfp M. smegmatis cells were nonspecifically labeled on their surface with the amine-reactive dye (AF-546), and unlabeled extensions were allowed to grow for 60 min. Representative image (E), and fluorescence quantification over the long axis of 22 cells (F) are shown (see also Fig. S2C). (Scale bar, 5 μm.)