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. 2022 Mar 29;13:1667. doi: 10.1038/s41467-022-29292-7

Fig. 5. Validation of resistance genes proposed by recommendation system.

Fig. 5

A Proliferation assays confirm osimertinib sensitivity of EGFR-mutant NSCLC cell lines PC-9, II-18 and HCC827. Data are presented as mean values +/– SD, n = 2 technical replicates, representative data of three biological replicates. B Resistance to osimertinib by target gene KO was measured in flow cytometry-based competition assays. Increase in percentage of KO cells compared to control cells was considered as resistance effect to osimertinib. C, D Confirmation of resistance to osimertinib in EGFR mutant NSCLC KO cell line models. Effect of KO of genes (EZH2, KCTD5, NF1 and PTEN) on osimertinib resistance was tested in II-18, PC-9 and HCC827 cell lines. Proliferation of cells in DMSO or osimertinib (50 nM) was tracked over 14 days and percentage of BFP+ KO cells was plotted. E Activation of MET drives resistance to osimertinib in EGFR mutant PC-9. Cells were engineered to express the dCas9-VP64 SAM CRISPR for MET activation. For C–E, use of two independent guide RNAs per gene were considered as biological replicates (n = 2), three technical replicates are visualized. Data are presented as mean values +/– SD. F Activation of WWTR1 drives resistance to osimertinib in EGFR mutant PC-9 cells as measured in 21 day clonogenic assays. PC-9 cells were engineered to express the dCas9-VP64 SAM CRISPR for WWTR1 activation. Data presented as mean values +/– SD, representative data of two biological replicates, each consisting of technical duplicates96.