Figure 7
From: Centrosomal kinase Nek2 cooperates with oncogenic pathways to promote metastasis
(a) Transient transfection of hNek2 in A549 cells led to upregulation of Akt pathway marker pS6, and strong deregulation of β-catenin levels. In mock-transfected cells β-catenin is at the cell–cell junctions but hNek2-transfected cells show strong reduction of β-catenin from the junctions. Note that some Nek2-transfected cells (asterisk) show increased β-catenin in and around the nucleus. (b) Western analysis of transiently transfected hNek2 in A549 cells shows upregulation of pAkt, pS6, pGSK3β, β-catenin, pJNK and Rac1 levels. Deregulated proteins indicated with asterisk. Constitutively active RET, hRETM918T, which is known to activate these pathways (pErk, pJNK, pAkt) is used as control. 40(c) Western analysis of transiently transfected hNek2 in HEK293T cells shows upregulation of pAkt, pS6, pGSK3β, β-catenin, pJNK and downregulation of RhoA. Relative levels of proteins between control and hNek2 overexpressing cells are indicated in red. (d) Nek2-dependent upregulation of β-catenin and pJNK, in HEK293T cells can be inhibited by addition of pelitinib or Akt inhibitor MK2206. Inhibitors were added at a concentration of 0.5 μM for 2 h, following 24 h of growth and transfection of cells.
