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Authors: Eva K. B. Pfannes, Christian Wegner, Matthias Theves, and Carsten Beta
We investigated the effect of the carbazole derivative Wiskostatin, a recently identified WASP inhibitor, on the mechanical stability and dynamics of motile Dictyostelium discoideum cells. The mechanical properties of cells were probed using a newly developed microfluidic device, the 'fluidic cell squeezer'. The core part of this device is a microfluidic channel that periodically changes its width. As cells travel through the series of fluidic apertures, they experience an alternating sequence of maxima and minima in flow speed and shear stresses. Depending on the mechanical stability of the cells, this lead to a deformation as they pass through the narrow parts of the channel. We observed that WASP-inhibited cells showed an increased deformability, similar to cells exposed to Latrunculin A, an inhibitor of actin polymerization. The total amount of F-actin however, remained constant in the Wiskostatin-treated cells. Further experiments with Wiskostatin showed an altered morphology and strongly reduced pseudopod formation. Cell-to-substrate adhesion was enhanced, and random motility as well as the chemotactic motion in gradients of cAMP were reduced. |
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