Extended Data Fig. 3: Expression, cell growth and cell morphology of FLAG-ess and PcomX-FLAG-ess ∆ess cells.
(a) Western immunoblot of whole-cell lysates of FLAG-ess, ∆ess and PcomX-FLAG-ess ∆ess cells, grown in the presence (3 µM) or absence (0 µM) of inducer (ComS), revealed with anti-FLAG antibody and anti-enolase antibody as a loading control. The expected band for FLAG-S Protein is indicated by the black triangle. (b) Representative phase contrast microscopy images of FLAG-ess, ∆ess and PcomX-FLAG-ess ∆ess cells, grown in the presence (3 µM) or absence (0 µM) of ComS inducer. Minicell-like compartments are highlighted by a white triangle. Scale bars, 2 µm. (c) Violin SuperPlots showing the distribution of the maximum cell width from three independent experiments, (n = 2000 cells for each strain). ***p < 0.001 and ns, not significant, p > 0.05. (d) Growth of FLAG-ess, ∆ess and PcomX-FLAG-ess ∆ess in C + Y medium at 37 °C. (e) Representative montage of images acquired by time-lapse microscopy, showing the formation of a minicell-like compartments in a ∆ess cell. The position of the ectopic division site is indicated by a white triangle. Time is given in min. Scale bar, 0.5 µm.
