VASP translocation to the cell periphery depends on S157 phosphorylation.
Wild-type endothelial cells (EC_VASP+/+) were incubated with
forskolin (5 μM) or buffer and analyzed using antibodies against
S157-P–VASP and total-VASP. (A) Western blots of cell lysates
normalized for GAPDH. (B-G) Immunofluorescence images of fixed and
permeabilized cells. The merge of total-VASP (red; B,E) and
S157-P–VASP (green; C,F) is given in yellow (D,G). Black
arrowheads indicate stress fibers, black arrows indicate focal adhesions, and
white arrowheads indicate plasma membranes. Images were taken with a
100× objective and are representative of a series of eight experiments.
Scale bar: 15 μm. (H) Comparison of VASP, Mena and EVL expression in
EC_VASP–/– and EC_VASP+/+ cells by western
blotting. The cell lysates are normalized for GAPDH.