Cell morphology and localization of FtsZ wild type and mutants E83Q and R85Q in cells with (30°C) and without (42°C) co-expression of FtsZ(WT) from pLAR9. Immunofluorescence of FtsZ (green) overlaid with the bright field image (gray) shows the localization of FtsZ in VIP5 cells (for strain information see Methods). Scale Bar, 5μm. Immunofluorescence was performed as described in Methods. Briefly, cells were grown until OD600 reached 0.5, cultures were split in half and incubated at 30°C and 42°C respectively for 4 h. Cells were harvested, fixed and immunofluorescent labeled with anti-FtsZ as first antibody and the second antibody conjugated with Alexa 488.