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Fig. 2 Effects of combination therapy with anlotinib and osimertinib on NSCLC cells in vitro. A CCK-8 assay was conducted to explore the proliferative capacities of (A) H1975, (B) PC9, (C) H1975-OR, and (D) PC9-OR cells, pre-treated with the NC, osimertinib monotherapy (1 µM), anlotinib monotherapy (3 µM), and a combination of anlotinib (3 µM) and osimertinib (1 µM) for 72 h. An MTT test was conducted to determine cell viability in (E) H1975-OR and (F) PC9-OR cells treated with different concentrations of osimertinib monotherapy or the combination of anlotinib (3 µM) and osimertinib. P values were calculated by two-way ANOVA analysis; ***, P<0.001; NS, not significant. OD, optical density; NC, negative control; Osi, osimertinib; Anl, anlotinib; Osi + Anl, osimertinib combined with anlotinib; NSCLC, non-small cell lung cancer; CCK-8, Cell Counting Kit-8; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide; ANOVA, analysis of variance.