Characterization of K-RAS mutant cell lines

K-RAS mutated cell lines A549, A427 and SK-LU1 were obtained from ATTC and grown in RPMI (Gibco) supplemented with 10 % FBS, 2 mM glutamine (Gibco) and penicillin/streptomycin (Gibco). 368T1 cells were a kind gift of Dr. Tyler Jacks (45 (link)). Pulm24 cells were derived from the PDX model and harbor a K-RAS G12C mutation. KP and KPE cell lines were isolated in house from respective mice by enzymatic digestion of lungs harvested following survival analysis for genotyping or 6 weeks after Ad.Cre mediated tumor induction for re-transplantation and K-ras activity assay. (45 (link)) To determine the density of viable cells we used a CASY counter (Roche).

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Moll H.P., Pranz K., Musteanu M., Grabner B., Hruschka N., Mohrherr J., Aigner P., Stiedl P., Brcic L., Laszlo V., Schramek D., Moriggl R., Eferl R., Moldvay J., Dezso K., Lopez-Casas P.P., Stoiber D., Hidalgo M., Penninger J., Sibilia M., Győrffy B., Barbacid M., Dome B., Popper H, & Casanova E. (2018). Afatinib restrains K-RAS driven lung tumorigenesis. Science translational medicine, 10(446), eaao2301.

Publication 2018

RPMI glutamine penicillin/streptomycin CASY counter

Assay Cell lines Cells Digestion Enzymatic Glutamine House mice Lungs Mutation Penicillin Streptomycin Transplantation Tumor

Corresponding Organization : Ludwig Boltzmann Institute for Cancer Research

Other organizations : Spanish National Cancer Research Centre, Medical University of Graz, Mount Sinai Hospital, University of Toronto, Austrian Academy of Sciences, Institute of Molecular Biotechnology, Lunenfeld-Tanenbaum Research Institute, University of Veterinary Medicine Vienna, Semmelweis University, Institute of Molecular Life Sciences, National Institute of Oncology

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Variable analysis

independent variables

Cell Lines (A549, A427, SK-LU1, 368T1, Pulm24, KP, KPE)

dependent variables

Density of viable cells

control variables

Growth media (RPMI supplemented with 10% FBS, 2mM glutamine, and penicillin/streptomycin)
Cell culture conditions

controls

Positive control: Not specified
Negative control: Not specified

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