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Figure 3. The effects of AZD1208 and osimertinib on RTKs and downstream components in EGFR-mutation positive NSCLC cells. A: in all cell lines, cell viability assays were performed to explore the effect of combined osimertinib and AZD1208 treatment. The obtained CoI was calculated based on the Chou and Talalay method, and values < 1, = 1 and > 1 indicate synergism, additive effect and antagonism, respectively. Experiments were performed in biological triplicates, and the average and standard deviations plus 95%CIs are shown; B: two EGFR-mutation positive NSCLC cell lines (H1975 and PC9) were treated with single osimertinib (0.024 µmol/L), single AZD1208 (5 µmol/L) or with the combination. Untreated cells received an equivalent dose of vehicle (DMSO). Cell lysates were used for immunoblotting and changes in RTKs and non-RTKs upon the different treatments were investigated in the two cell lines. Experiments were performed in biological triplicates with similar results, and a representative blot is shown. RTK: receptor tyrosine kinase; EGFR: epidermal growth factor receptor; NSCLC: non-small cell lung cancer; CoI: combination index; 95%CI: 95% confidence interval; DMSO: dimethylsulfoxide; CDCP1: CUB domain -containing protein 1; YAP1: yes-associated protein 1; STAT3: signal transducer and activator of transcription 3