Cytoskeletal Interactions at the Nuclear Envelope Mediated by Nesprins

Figure 2

N-terminal spectrin repeats of Nesprin-1 undergo self-associations. (a) Schematic illustration of Nesprin-1-165 constructs used as GST and GFP fusion proteins. Numbers indicate the location of amino acids. (b, c) COS7 cells were transfected with GFP-tagged ABD (aa 1–286), and spectrin repeats of Nesprin-1-165 (aa 573–858, 859–1144, and 1145–1431) (b) and full-length Nesprin-1-165 (c). The cells were lysed in RIPA-buffer and equal amounts of each lysate were incubated with either immobilized GST-fused aa 1–286, 573–858, 859–1144, and 1145–1431 or GST alone for control shown in (d) and (e). The pull down samples were subjected to SDS-PAGE followed by western blot analysis using GFP antibody mAb K3-184. (d, e) The GST-fusion proteins are revealed by Coomassie Blue staining.