WASp^EMM reconstitutes and localizes to structurally characteristic podosomes.^WASBMDCs were plated onto fibronectin-coated glass coverslips on day 7 of culture. After 4 hours of adherence, cells were fixed and immunostained for EGFP (rabbit anti-EGFP and anti–rabbit Alexa Fluor 488), f-actin (Alexa Fluor 568 phalloidin), and vinculin (mouse anti-vinculin and goat anti–mouse Cy5). Images were taken using a Zeiss LSM710 laser scanning spectral confocal microscope and a 63× Plan-Apochromat NA 1.4 WD 190-mm oil immersion lens and processed using ImageJ Version 1.45d software. Images represent single Z stack slice through the adhesive surface of the cell, and adjustments were made to brightness, contrast, and γ only. For whole-cell images, actin is shown in white, vinculin in yellow, and EGFP-WASp in green. To aid the visualization of colocalization, in the podosomes detail panels, the colors are changed to actin in red, vinculin in blue, and EGFP-WASp in green. The bottom right panel for each construct represents a merged image of all 3 channels.