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. Author manuscript; available in PMC: 2011 Apr 1.
Published in final edited form as: Exp Mol Pathol. 2009 Dec 23;88(2):225–233. doi: 10.1016/j.yexmp.2009.12.004

Fig. 3.

Fig. 3

Nek2-Cdc20 interaction. (A) Co-immunoprecipitation analysis of Mad2-Cdc20 interaction. 293T cells were transfected with GFP-Cdc20 in combination with FLAG-Nek2A or the empty control plasmid. Upper panel: IP with anti-FLAG antibody; immunoblotting with anti-GFP antibody. Lower panel: Western Blot using anti-GFP antibody. (B) Reciprocal co-IP. 293T cells were transfected with the plasmids, as indicated. Upper panel: IP with anti-GFP antibody; Western blot using anti-FLAG antibody. Lower panel: Western blot using anti-FLAG antibody. (C) 293T cell lysate was subjected to IP using anti-Nek2 antibody (lane 3) or control mouse IgG (lane 2) followed by Western blot analysis using anti-Cdc20 antibody. The endogenous Cdc20 level in the cell lysate was also examined (lane 1). (D) The Nek2 region involved in association with Cdc20. 293T cells were transfected with the plasmids, as indicated. Top panel: IP with anti-FLAG antibody; Western blot (WB) using anti-GFP antibody. Middle panel: IP and WB using anti-FLAG antibody. Bottom panel: Western blot analysis of cell lysate using anti-GFP antibody.