Gigantosomes are coated with clathrin. HeLa cells stably expressing EGFP-CLC (green) and transfected with mRFP–EEA-1 (red) were analyzed by live 3-dimensional confocal capture 10 minutes after sublytic rPFN treatment. In the presence of rPFN, gigantosomes stained with EEA-1 (red), colocalize with clathrin (green) as indicated. One representative optical section obtained from sequential 0.1-μm optical sections (supplemental Videos 3-4) is shown for each condition. Most of the gigantosomes (red) are coated with clathrin (green). Data are representative of 5 independent experiments. Color bars and associated numbers indicate fluorescence intensity levels. Scale bars represent 10 μm. Dashed lines indicate plasma membrane.