The cell line HCC827 was purchased from ATCC. An erlotinib-resistant HCC827 cell line (HCC827ER) was generated as previously described [21 (link)]. Further, HCC827ER clones were established using minimal dilution [21 (link)]. All cells were maintained in RPMI 1640 media supplemented with 1% Penicillin-Streptomycin (Gibco), 1% Hepes 1 M buffer solution (Gibco), 1% Sodium Pyruvate (Gibco), 10% Fetal Bovine Serum (Gibco), and 1% 250 μg/mL Amphotericin B solution (Sigma-Aldrich). Further, the resistant cell line and clones were maintained in 5 μM erlotinib (Selleckchem).
For all mRNA and flow cytometry experiments cells were plated in 6-well plates and incubated for 24 h (300,000 cells/well). After incubation the cells were treated with drug and incubated for 72 h prior to harvest.
The inhibitors erlotinib, crizotinib, and SCH772984 were all purchased from Selleckchem.
For all mRNA and flow cytometry experiments cells were plated in 6-well plates and incubated for 24 h (300,000 cells/well). After incubation the cells were treated with drug and incubated for 72 h prior to harvest.
The inhibitors erlotinib, crizotinib, and SCH772984 were all purchased from Selleckchem.
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