NMII is associated with the bleb cytoskeleton in different types of blebs. Immunogold PREM of blebbing HT1080 cells treated with 200 µM CK-666. (A and E) Anaglyph stereo images of blebs at the cell periphery. (B and C) Enlarged 2D images of an early-stage bleb (B) and a late-stage bleb (C) from A. Immunogold particles are pseudocolored yellow. (D) A small actin filament bundle in the bleb interior from the boxed region in A and B shown in 2D without pseudocolor. As can be appreciated from A, this NMII-associated bundle ascends from its tip (at the top of the image) toward the base (at the bottom of the image), while NMII immunogold particles associated with the bundle are located inside the bleb. (F) Two middle-stage blebs with cortical enrichment of the cytoskeleton enlarged from E shown as a 2D image with NMII immunogold pseudolcolored yellow. Scale bars: 500 nm (A and E), 200 nm (B and F), and 100 nm (C and D).