Paxillin Ser-85 phosphorylation affects cell adhesion signaling activity, FA formation, and actin organization.
A, cells were transfected with HA3-paxillin (WT, S85A, or S85D) for 36 h before being kept in suspension (Sus) or reseeded onto collagen I (Coll; 10 μg/ml)-precoated culture dishes for 30 min. Whole cell lysates were prepared for standard Western blots for the indicated molecules. pS85Pax, phospho-Ser-85 paxillin. B, HeLa cells were transfected with HA3-paxillin (WT, S85A, S85D, or S85E) for 36 h before replating on collagen I (10 μg/ml)-precoated coverslips for 30 min. The cells were stained for actin using phalloidin and immunostained for HA3-paxillin using anti-HA antibody. Scale bar = 10 μm. The data shown represent three independent experiments.