Figure 4.

Vinculin and zyxin associate with different structures in podosomes. (A–D) DCs were seeded on glass coverslips and after VPM preparation, cells were fixed and stained for vinculin (green, B), zyxin (magenta, C) and actin (cyan, Supplementary Figure 4). Overlay between vinculin and zyxin is shown in white (D). After CPD, DCs were imaged by SEM (gray, A). Shown is a representative podosome with radiating filaments that are clearly observed in the SEM image. Arrows indicate areas enriched for vinculin or zyxin, which overlap with the radiating actin filaments (also indicated by arrows). Scale bar = 500 nm. (E–F) Radial fluorescence profile analysis of vinculin (E) and zyxin (F) at each of the four z-sections depicted in (B–D). Shown is the average ± SEM (n = 312 podosomes, 5 cells). (G) Quantification of the localization in z of zyxin, vinculin, core actin and network actin in podosomes. The z-sections shown in (B–D) are represented by the dashed lines in the graph. Shown is the average ± SEM (n = 312 podosomes, 5 cells).