Figure 5. An AAK1 interaction network promotes EMP and therapy resistance in HCC and exhibits characteristics of a proliferation suppressor.
(A) Kinobead/LC-MS soluble competition experiments using 1 μM of the selective AAK1 inhibitor LB-935509 show that the PPI network is centered on AAK1, not BMP2K.
(B) Immunoblotting of EMP markers in SNU387 and FOCUS AAK1 network RNAi lines indicated alterations in EMP state.
(C) Heatmap showing EMP-associated receptor kinases that change in abundance in response to AAK1 PPI network RNAi in mesenchymal-like HCC cells. Kinase abundance differences were determined by kinobead/LC-MS profiling.
(D) Drug screen results demonstrating that AAK1 network RNAi lines are up to 18-fold more sensitive to the cell cycle checkpoint kinase (CHEK1) inhibitors AZD-7762 and CHIR-124.
(E) Kinobead/LC-MS profiling of SKHep1 and SNU387 lines shows that AAK1 and REPS1 RNAi causes upregulation of cell cycle-related kinases and their activating PPIs specifically in lines with increased sensitivity to CHEK1 inhibitors.
(F) Immunoblotting confirmed activation of the cell cycle specifically in AAK1 and REPS1 RNAi lines, suggesting that they act as proliferation suppressors in mesenchymal-like cells.