Fig. 1. Main features of invadopodia and effect of BB-94 treatment. (A) Triple staining of A375MM cells grown on crosslinked FITC-conjugated matrix for 16 hours and then fixed and stained with anti-cortactin (A) and Alexa Fluor 633-phalloidin (B), or anti-dynamin 2 (E) and anti-phosphotyrosine (F). All these proteins colocalized precisely to ECM degradation patches (C and G, arrows). (B) Anti-cortactin (A,E) and Alexa Fluor 633-phalloidin (B,F) labeling of cells grown for 16 hours on fluorescent matrix (C,G) in the continuous presence (+BB94) (A-D) and 3 hours after washout (–BB94) of the inhibitor (E-H). Scale bars: 10 µm.