Fig. 5. (A-C) Immunofluorescent images showing the sub-cellular localization of nesprin-2 isoforms using N2, N3 and CH3 antibodies co-localized with lamin, in human VSMCs. All three antibodies localize to domains in the nucleus with N2 and CH3 showing nuclear rim staining and N2 and N3 abundant around nucleoli (identified by domain-specific antibody; not shown). Cytoplasmic staining was also observed using all three antibodies and this is shown in detail using human skin fibroblasts. (D-F) In the cytoplasm of skin fibroblasts, nesprin-2 is diffusely distributed in a reticular pattern and is localized to lamellipodia (arrow in D), to focal adhesions (arrows in E) and to filopodia (arrow in F; red stain shows ß-tubulin). All stainings were competed completely by excess peptide (not shown).