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. Author manuscript; available in PMC: 2011 Dec 23.
Published in final edited form as: Cell. 2010 Dec 23;143(7):1097–1109. doi: 10.1016/j.cell.2010.11.038

Figure 4.

Figure 4

LpoA and LpoB localize as distinct foci in the lateral wall and at constriction sites of dividing cells. E. coli wildtype (TB28) (A) and its lpoA− derivative (C) were immunolabeled with antibodies against LpoA. E. coli wildtype (BW25113) (B) and its lpoB− derivative (D) were immunolabeled with affinity-purified antibodies against LpoB. The immunolocalization procedure does not affect the cell membrane (Fig. S5A) or the size/shape of the cells (Supplemental Experimental Procedures). The left side of each dual panel shows the phase contrast image and the right side the corresponding fluorescence image. The scale bar equals 5 μm. Arrows in panels A & B depict LpoA and LpoB foci for cells engaged in septation. Panels E–H show the average LpoA (E & G) or LpoB (F & H) fluorescence intensity profiles of >1000 individual cells per strain plotted against the relative position along the length axis of the cell. The populations of cells were split into longer cells (1/3 of the population), enriched in dividing cells (E & F) and shorter cells (2/3 of the population), including only few dividing cells (G & H). For panels E–H: black lines: wildtype cells; red lines: lpoA− cells; blue lines: lpoB− cells; green lines: mrcA− cells (lacking PBP1A) and purple lines: mrcB− cells (lacking PBP1B). The grey line in panels E & G are from a general membrane staining using BODIPY 558/568 C12. LpoB localizes late in the cell cycle to midcell (Fig. S5B). Midcell localization of LpoB depends on the presence of FtsZ, PBP3 and ongoing septal PG synthesis (Fig. S6).