Fig 6. Effect of cell division inhibitors on localization of (A) Cls_6xH (labeled Cls) and (B) MreB_6xH (labeled MreB).

Cells were infected with transformants carrying anhydrotetracycline (aTc) inducible vectors encoding the indicated constructs. Expression of Cls_6xH or MreB_6xH was induced at 16 hpi, and cells were fixed and processed for immunofluorescence at 20 hpi. To visualize peptidoglycan (PG), cells were incubated with EDA-DA during the infection and processed using click-iT reagents as described in the Materials and Methods. To assess effects of disrupting the central coordinator of chlamydial cell division, MreB, the antibiotic A22 was added at 18hpi whereas peptidoglycan synthesis was disrupted using D-cycloserine (DCS) added at 4hpi. Note the continued presence of Cls_6xH at a discrete site on the bacteria under all conditions tested. The insets within the upper row images are a zoomed-in view of the boxed regions in the larger image. Images were acquired on a Zeiss AxioImager.Z2 equipped with an Apotome2 using a 100X lens objective. Images are representative of at least three independent experiments. Scalebar = 2 μm.