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. 2011 May;10(5):662–671. doi: 10.1128/EC.00221-10

Fig. 2.

Fig. 2.

Evidence for functional significance of the SadA carboxy-terminal tail. (A) Micrographs of sadA null cells expressing either a wild-type SadA or a SadA carboxy-terminal deletion construct (SadA ΔTail+3-GFP construct) imaged with a confocal microscope. Increased levels of GFP signal were generally observed in cytoplasmic intracellular vesicles compared to the plasma membrane compartment. The right panel is a micrograph of cells imaged using the agar overlay technique, and the left panel shows cells without overlay. Bar, 10 μm. (B) Adhesion maintenance assay of AX3 wild-type and sadA null cells expressing the SadA ΔTail+3-GFP construct. Whereas AX3 cells (•) exhibited a sheer force-dependent decrease in substrate adhesion, the SadA ΔTail+3-GFP-expressing cells (▪) had minimal adhesion at all speeds tested. (C) Adhesion maintenance of AX3 cells (•) and AX3 cells overexpressing the SadA carboxy-terminal tail (▪). AX3 cells overexpressing the SadA tail were less resistant to increased sheer stress than AX3 wild-type cells. Error bars represent the standard deviations from triplicate or quadruplicate samples. (D) Triton solubility assay of AX3 and sadA− plus pTXSadAΔtail+3-GFP. The experiment was conducted as described for Fig. 1, but only under control (untreated) conditions. Whereas wild-type SadA partitioned primarily into the Triton X-100-insoluble fraction, SadAΔTail+3 partitioned, almost equally, between the insoluble and soluble fractions.