Fig. 5

AXL inhibitor prevented the emergence of drug-tolerant cells to osimertinib. a Drug-tolerant (DT) cells previously treated with 3 μmol/L osimertinib for 9 days were treated with the indicated concentrations of osimertinib for 72 h and their viability was assessed using MTT assays. b PC-9 parental cells and DT cells generated by treatment with 1 or 3 μmol/L osimertinib were lysed and the indicated proteins were detected by western blotting. c PC-9 parental cells and DT cells generated by treatment with 3 μmol/L osimertinib for 9 days were lysed and the indicated proteins were detected by western blotting with immunoprecipitation of the indicated proteins. d PC-9, HCC4011 parental cells, and DT cells generated by treatment with 3 μmol/L osimertinib were treated with 1 μmol/L of NPS1034, OSI906, or a combination of these agents for 72 h and the cell viability was assessed using MTT assays. e The indicated cells were incubated with osimertinib (3 μmol/L) with or without NPS1034 (1 μmol/L) for 1 h. The cells were lysed and the indicated proteins were detected by western blotting. f Cells were treated with DMSO, 100 nmol/L osimertinib, 1 μmol/L NPS1034, or a combination of 100 nmol/L osimertinib and 1 μmol/L NPS1034 for 3 weeks with the drugs replenished every 72 h. The plates were stained with crystal violet and visually examined. A plate representative of three independent experiments is shown