Inhibition of Nek2 by Small Molecules Affects Proteasome Activity
Figure 1
Nek2 overexpression causes HeLa cells to become resistant to bortezomib. (a) The Nek2 gene was cloned into a GFP expression vector as described in Materials and Methods Section. HeLa cells were then transfected with either the Nek2-GFP plasmid or GFP expression vector alone. Anti-NEK2 antibody was used to confirm NEK2 overexpression as determined by Western blot. (b) Nek2 overexpression increased the level of phosphorylated PP1-α in the two surviving Nek2 transfected clones. (c) Nek2-GFP transfected HeLa cells were resistant to bortezomib treatment compared to GFP-transfected clones. Bortezomib was used to treat HeLa cells with the concentration range from 100 nM to 0.03 nM. Within this range, at any given concentration of bortezomib, Nek2-transfected clones yielded higher cell viability than GFP-transfected clones.
