Fig. 1.

Swiprosin-1 colocalizes with F-actin and modulates lamellipodia formation. a Anti-Swiprosin-1 antibody (Sw1-R Ab) is a rabbit polyclonal antibody raised against recombinant human Swiprosin-1 protein. The antibody was compared with goat anti-Swiprosin-1 Ab (anti-Sw1-G) in IB experiments with B16F10 cells (upper left panel). Lysates were obtained from B16F10 cells transfected with empty GFP vector or GFP-Swiprosin-1 for 24 h. GFP-Swiprosin-1 and transfection efficiency were detected with anti-Sw1-R, anti-Sw1-G, and anti-GFP antibodies (upper right panel). GFP signals merged with the anti-Sw1-R antibody staining (lower panel). b B16F10 cells were treated with or without 100 ng/ml EGF for 2 min, and stained with anti-Sw1-R antibody and phalloidin-594 to visualize F-actin. c B16F10 cells were transfected with empty GFP vector or GFP-Swiprosin-1 for 24 h. Next, cells were stained with Alexa-594-phalloidin. Merged images are shown in yellow. Scale bar 10 μm. Lamellipodia formation was determined with actin staining among GFP-expressing cells. (n > 30 cells, *P < 0.005). d B16F10 cells were transfected with psi-RNA-GFP-control or psi-RNA-GFP-Swiprosin-1 for 24 h, and 100 ng/ml EGF administered for 2 min for lamellipodia induction. The transfected cells were stained with Alexa-594-phalloidin, and the lamellipodia formation was analyzed in GFP-expressing cells (*P < 0.05, **P < 0.01)