EM of cells recovering for 15 min after washout of 100 µM blebbistatin.

<p>(A–C) Correlative fluorescence (A) and EM (B,C) of REF52 cell stained with phalloidin (A, red) and vinculin antibody (cyan) and additionally labeled with NMII immunogold (C, yellow). (A, B) Same cell region shown by fluorescence microscopy (A) and by EM overlaid with vinculin immunofluorescence (B). Focal complexes are located at the bases of lamellipodia (upper left and lower right) and in the body of lamella. Mature focal adhesions (center) colocalize with actin bundles (box). (C) Enlarged box from B. NMII immunogold particles form multiple elongated clusters aligned with actin bundles. (D,E) EM of gelsolin-treated cytoskeleton. Arrowheads point to longitudinal chains of NMII filaments at cell periphery (D) or cell interior (E). Scale bars, 5 µm (A,B) and 1 µm (C–E).