Fig. 5: The C-terminal domain of DivIC is essential for function and septal localization.
a A hypothetical topological model of the S. aureus DivIC protein. b Viability test of ΔdivIC carrying the empty vector (SJF5544) or expressing GFP-DivIC (SJF5473), GFP-DivICΔA100-K130 (SJF5553) and GFP-DivICΔE70-K130 (SJF5554) in the presence and absence of IPTG. Data represents the mean and standard deviation of at least three independent experiments. P-values were determined by two-tailed unpaired t-test (****P < 0.0001). c Western blot of whole cell lysates from the ΔdivIC strain carrying the empty vector or expressing GFP-DivIC, GFP-DivICΔA100-K130, or GFP-DivICΔE70-K130. Cells were grown without IPTG for 2 h. Anti-GFP antibodies were used for detection. d Localisation of GFP-DivIC, GFP-DivICΔA100-K130 and GFP-DivICΔE70-K130 in ΔdivIC cells grown without IPTG for 2 h and incubated with HADA for 30 min to label PG. e FR from GFP-DivIC, and GFP-DivICΔE70-K130 signals (from data based on d). f Localisation of GFP-DivIC in SH1000 (SJF5424) and ΔdivIB (SJF5427) cells growing with and without IPTG for 2 h and incubated with HADA for 30 min to label PG. g FR from GFP-DivIC (from data based on f). e and g Each circle indicates a single cell, lines are mean of an n = 100. P-values were determined by Mann–Whitney U-tests (*P = 0.0238, ****P < 0.0001). All images are average intensity projections of z stacks of cells with incomplete septa. Scale bars, 2 μm. FR are the product of the fluorescence signal at the septum divided by the signal at the cell periphery in cells with incomplete septa. Data is representative of two independent experiments.
