Figure 1

EGFR-L858R expression and lung cancer cell invasion ability.

(a) Over-expression of EGFR and EGFR-L858R in lung adenocarcinoma H1299 cells was evaluated by Western blotting. H1299-EGFR-WT and H1299-EGFR-L858R cells were stably transfected with EGFR-WT and EGFR-L858R, respectively; H1299-mock was a mock-transfected cell line used as a control. EGFR-L858R expression in H1299-EGFR-L858R cells was knocked down using an EGFR-L858R–specific siRNA. The full size blots were shown in the Supplementary Figure S2 and band of interest is indicated with an arrow. (b) EGFR-L858R expression was quantified by real-time RT-PCR (*P < 0.001). (c) Using an EGFR-L858R–specific siRNA, EGFR-L858R expression was knocked down in H1299-EGFR-L858R cells. EGFR protein was evaluated by Western blotting. The full size blots were shown in the Supplementary Figure S2 and band of interest is indicated with an arrow. (d) The invasiveness of cells constitutively expressing EGFR-WT or EGFR-L858R in H1299 cells was evaluated using a modified Boyden chamber assay (*P < 0.01). (e) CL1-0 cells were transiently transfected with pCDNA4-EGFR-WT and pCDNA4-EGFR-L858R plasmids by using Lipofectamine method. The expression of EGFR-WT and EGFR-L858R in lung adenocarcinoma CL1-0 cells was evaluated by Western blotting. The full size blots were shown in the Supplementary Figure S2 and band of interest is indicated with an arrow. (f) The invasiveness of cells expressing EGFR-WT or EGFR-L858R in CL1-0 transfectants was evaluated using a modified Boyden chamber assay (*P < 0.01). (g) The invasion ability of H1299-EGFR-WT and H1299-EGFR-L858R cells was determined using a modified Boyden chamber after siRNA-mediated knockdown of EGFR-L858R (*P < 0.01). si-Scramble, cells transfected with scrambled siRNA control; si-EGFR-L858R, cells transfected with EGFR-L858R–specific siRNA. Data are presented as means ± SD of three independent experiments. P-values (two-sided) were determined using Student’s t-test.