Fig. 6.

Effects of crizotinib on ALK/MET phosphorylation, AKT and ERK activation and cell growth through IGF-1 signalling in RH4 and RH30 cell lines. a Immunoprecipitation and western blot showing expression of p-ALK, total ALK and p-MET and total MET in RH4 and RH30 cells treated with or without 1.5 μM crizotinib (Crz) for 24 h. Control cells (Ctrl) were treated with DMSO alone. b Western blotting analysis of phosphorylation levels of AKT and ERK proteins and downstream molecules GSKβ and P70S6K in RH4 and RH30 cells treated with or without 1.5 μM crizotinib (Crz) for 24 h. Control cells (Ctrl) were treated with DMSO alone. c Western blotting analysis of phosphorylation levels of IGF1R and AKT proteins in IGF1-treated cells with or without crizotinib. RH4 and RH30 cells were starved in serum deficient medium for 24 h prior to induce with IGF1 (50 ng/ml) for 30 min. Crizotinib (1.5 μM) was added 4 h before ligand stimulation. d MTT proliferation assay in ARMS cells grown in the presence or absence of IGF1 (50 ng/ml) and crizotinib (1.5 μM) for 48 h. Bars represent the mean values for nine replicate wells ± SD and are representative of three independent experiments