Figure 1.

FYCO1-positive LE/Lys localize to invadopodia. (A) MDA-MB-231 cells were grown on coverslips, stained with antibodies against TKS5 and cortactin, and analyzed by confocal microscopy. Phalloidin/Alexa Fluor 647 was used to detect F-actin. A section from a confocal z-stack shows invadopodia. Orthographic sections show invadopodia on the ventral side of the cell. (B) MDA-MB-231 cells were grown on coverslips coated with Oregon Green gelatin for 4 h, stained with anti-TKS5 and phalloidin/Alexa Fluor 647 (actin) to visualize invadopodia, and analyzed by confocal microscopy. A section from a confocal z-stack shows invadopodia correlating with degraded gelatin (black areas). 3D view and orthographic sections of the same cell are shown. (C) Colocalization of FYCO1 with LE/Lys markers in MDA-MB-231 cells. Cells were grown on coverslips; stained with antibodies against FYCO1, RAB7, or LAMP1; and analyzed by confocal microscopy. (D) MDA-MB-231 cells were grown on coverslips coated with Oregon Green gelatin for 4 h, stained with antibodies against TKS5 and FYCO1, and analyzed by superresolution microscopy (Airyscan). Z-stacks and Imaris surface 3D renderings from two independent cells show FYCO1-positive LE/Lys in close apposition to TKS5-positive invadopodia correlating with degraded gelatin. Data are representative of at least 16 captures.