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. Author manuscript; available in PMC: 2016 Aug 1.
Published in final edited form as: Nat Med. 2016 Feb 1;22(3):262–269. doi: 10.1038/nm.4040

Figure 1. Variable sensitivity of resistant T790M PC9 cell lines to EGFR inhibition.

Figure 1

(a) PC9 parental cells were cultured in escalating concentrations of gefitinib as tolerated until fully resistant, which was defined as lack of inhibitory effect of drug on cell proliferation.

(b) PC9-GR3, PC9-GR2 and parental PC9 cells were cultured with 1 μM gefitinib (G), WZ4002 (W) or vehicle (V) for 72 hours and apoptosis was determined by annexin staining (mean and s.e.m. of four independent experiments*P < 0.05, two-tailed t-test.).

(c) PC9-GR3, PC9-GR2 and parental PC9 cells were treated with 1 μM gefitinib (GEF), WZ4002 (WZ) or vehicle (VEH) and cell proliferation was determined by CellTiter-Glo assay at indicated time points (mean and s.e.m. of 4 independent experiments). The dotted line indicates relative cell number at time of drug addition.

(d) Mice bearing PC9-GR2 or PC9-GR3 subcutaneous xenograft tumors were treated with 50 mg/kg/day WZ4002. (PC9-GR2 - control (N=8), WZ (N=8); PC9-GR3 - control (N=8), WZ (N=8)). Tumors were measured with electronic calipers and % tumor response was calculated as the percentage change in tumor volume (V = 0.52 × L × W2) relative to the start of drug treatment (mean and s.e.m.). P < 0.01 (*) comparing WZ treatment arms at indicated time points by multiple t-tests with Sidak-Bonferroni multiple comparison.