FIG. 5.

LytF-3xFLAG localization in MinCD-depleted cells. B. subtilis WE1EMCp (wprA epr lytF-3xFLAG Pspac-minCD) was first grown in LB medium with 0.2 mM IPTG at 37°C to an OD₆₀₀ of 0.5 to 0.7. Cells were harvested and inoculated at an OD₆₀₀ of 0.08 into 5 ml of fresh LB medium without IPTG. After 2 h of incubation, cells were harvested and fixed as described in Materials and Methods. Cell wall staining (red) was performed with WGA-TMR at a final concentration of 1 μg/ml. The overlay was obtained by superimposing a DAPI-stained fluorescence image (blue), an FITC-stained image (green), and a WGA-TMR-stained image (red). The exposure times were 0.1 s for phase-contrast microscopy, 0.005 s for DAPI, 0.1 s for FITC, and 0.05 s for rhodamine. Bars = 5 μm.