Fig. 1.

SadA associates with the F-actin cytoskeleton. AX3 cells grown under adherent conditions received fresh medium (control) or were treated with DMSO or 5 μM latrunculin A (LatA). Cells lysed with 1% Triton X-100 (L) were subjected to low-speed centrifugation, divided into pellet (P) and supernatant (S) samples, and fractionated by SDS-PAGE followed by Western blotting. Membranes were probed with a polyclonal SadA antibody followed by a monoclonal anti-actin antibody. In control and DMSO-treated cells, SadA partitioned into the Triton-insoluble pellet, and actin was divided between the pellet and supernatant fractions. When cells were treated with latrunculin A, both SadA and actin were observed to fractionate completely into the Triton X-100-soluble fraction.