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. Author manuscript; available in PMC: 2014 Jul 21.
Published in final edited form as: Mol Microbiol. 2012 Jul 25;85(6):1090–1104. doi: 10.1111/j.1365-2958.2012.08159.x

Fig. 2. Phenotypes of MbiA overexpression.

Fig. 2

A. Growth curves of Caulobacter cells carrying pXyl::mbiA grown in PYE supplemented with either xylose (MbiA OE) or glucose (No OE). Normalized results of two experiments are presented.

B. Proportion of propidium iodide-positive cells carrying pXyl::mbiA after 7 h of growth in PYE with glucose (No OE) or xylose (MbiA OE).

C. Phase contrast images of cells carrying pXyl::mbiA and grown in PYE with xylose (MbiA OE) or glucose (No OE) for 4 h or overnight (around 24 h).

D. Cells carrying pXyl::mbiA grown in PYE with xylose for 8 h were placed on PYE pads with glucose to repress MbiA overexpression and imaged at indicated times after the transfer. Arrowhead indicates the progeny of the recovering cell that fails to divide. The panel at the right shows an isogenic strain grown in PYE with glucose and imaged at the 0 h time point.

E. Phase contrast, GFP-MreB fluorescence, and overlay images of swarmer cells immediately after synchronization and chemical fixation. LS3814 cells carrying (MbiA OE) or not carrying (No OE) pXyl::MbiA were grown in PYE with xylose. The plots at the right represent the average fluorescence intensity (in arbitrary units) at each position along the longitudinal axis (in % cell length) for the entire population of swarmers. n = 1126 for MbiA OE and n = 859 for No OE.

F. Immunoblot with anti-MreB and anti-DivJ antibodies of cells expressing either wild-type levels, increased levels, or no MbiA. Lane 1: CB15N; Lane 2: ΔmbiA (ZG708); Lane 3: MT196 with xylose; Lane 4: MT196 with glucose; Lane 5: MT196 pXyl::mbiA with xylose; Lane 6: MT196 pXyl::mbiA with glucose; Lane 7: CB15N pBV-mbiA (vanillate-inducible overexpression vector) with vanillate; Lane 8: CB15N pBV-mbiA without vanillate; Lane 9: CB15N pBVMCS-4 (empty vector) with vanillate. All cells were grown in PYE for 5 h.

G. Colocalization of mCherry-MreB and FtsZ-CFP in ZG695 swarmers carrying (MbiA OE) or not carrying (No OE) pXyl::MbiA. Cells were grown in PYE with xylose, inducing FtsZ-CFP for 1 h with 50 µM vanillate, and the images were taken immediately after synchronization and chemical fixation. Phase contrast, mCherry-MreB fluorescence, FtsZ-CFP fluorescence, and overlay images are provided. All scale bars are 2 µm.