Localization of cross-linking proteins in fibroblast cytoskeleton. Fluorescence microscopy and corresponding intensity profiles of Xenopusfibroblast lamellipodia double stained with TRITC-phalloidin (red) and either p21 (Arp 2/3) antibodies (green). The protein/actin ratio at the leading edge of the lamellipodium is high for Arp2/3 complex. Immuno-EM of the cell edge (CIL 34925) or interior (CIL 34928) of cytochalasin D-treated Xenopus fibroblasts stained with p21 primary antibody and 10-nm gold-conjugated secondary antibody reveal Arp2/3 complex at Y-junctions. Image corresponds to Figure 5a and a' from J Cell Biol. 1999 May 31;145(5):1009-26.
Procedures for detergent extraction, immunostaining, S1 decoration, light, and EM were described previously (Svitkina et al., 1995, 1996, 1997;Verkhovsky et al., 1995; Svitkina and Borisy, 1998).
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