Fig. 3: Characterization of EGFR variants of unknown significance (VUS) in lung cancer models of EGFR addiction.
From: Comprehensive mutational scanning of EGFR reveals TKI sensitivities of extracellular domain mutants
a EGFR domain schematic showing tested variants of unknown significance (VUS). b, c PC9, HCC4006, or HCC827 cell lines expressing either LacZ, EGFR WT, EGFR T790M, EGFR extracellular domain variants (R222C, S229C, A237Y, T302H, C311R, S447F, C595G, or P644S) (b), or EGFR intracellular domain variants (T725M, V769M, H773R, or V774M) (c) after 144 h treatment with increasing doses of erlotinib (normalized to vehicle control). A representative experiment is shown, remaining biological replicates are located in Source Data (N = 3). Data are presented as mean values +/- SD (b, c). d-f Colony formation with PC9 EGFR mutants and controls as in (b) and (c) after 10 d (PC9) or 14 d (HCC4006 and HCC827) of treatment with either vehicle (DMSO), 100 nM erlotinib (HCC4006 or HCC827), or 200 nM erlotinib (PC9). A representative experiment is shown (N = 3). g–l Representative immunoblots displaying the effect of either 200 nM (PC9) or 100 nM (HCC4006) erlotinib after 24 h treatment of cell lines expressing either LacZ, EGFR WT, EGFR T790M, EGFR extracellular domain variants (R222C, S229C, A237Y, T302H, C311R, S447F, C595G, or P644S) (g, h, i), or EGFR intracellular domain variants (T725M, V769M, H773R, and V774M) (j, k, l) on the levels of both phosphorylated EGFR and ERK and total EGFR and ERK. β-actin immunoblotting was used to determine equivalent loading. Data are presented as mean values ± SEM of biological replicates (PC9, N = 4; HCC4006, N = 3) (i, l).
