1. Fig. 4. Functional annotation of VUS in EGFR.
Functional annotation of VUS in EGFR. (A) Venn diagram revealing the numbers of oncogenic EGFR mutants seen in 3T3 cells using a focus-formation assay (3T3 FF), 3T3 cells by the MANO method (3T3 MANO), Ba/F3 cells using the MANO method (Ba/F3 MANO), or BA/F3 cells individually examined for IL-3–independent growth (Ba/F3 individual). The method for the evaluation of the transforming activity in each assay is described in detail in the legend for fig. S6. (B) Ba/F3 cells expressing the L858R, A839T, or T790M_C797S_Cis mutants of EGFR were treated with the indicated concentrations of gefitinib, erlotinib, afatinib, or osimertinib for 72 hours. Viable cells (%) were measured using the alamarBlue cell viability assay, and the results are shown as the means of five independent experiments. (C) Ba/F3 cells expressing 86 EGFR mutants (indicated at top) were treated with either DMSO or EGFR TKIs (gefitinib, erlotinib, afatinib, osimertinib, or rociletinib) at , , 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, or 10 μM, and the relative viability of TKI-treated cells relative to the corresponding DMSO-treated cells is color-coded according to the scheme indicated at the top left. Missense, deletion, and insertion mutations are shown in green, brown, and red, respectively.
Shinji Kohsaka et al., Sci Transl Med 2017;9:eaan6566
Published by AAAS