Dialysed fbs

All the human cell lines (Hs766T, BxPc3, Patu8988T and Patu8902) used in this study were purchased from ATCC, used below passage 25 and continuously cultured in 100 U/ml penicillin and 100 U/ml streptomycin. The cell lines were authenticated by short tandem repeat (STR) profiling at the Institute for Applied Cancer Sciences, MD Anderson Cancer Center. The Patu8988T, Hs766T and Patu8902 cell lines were routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS (Invitrogen). BxPc3 cell lines were routinely cultured in Roswell Park Memorial Institute (RPMI) 1640 (Invitrogen) with 10% FBS. Primary mouse cell lines were established in the laboratory (AK-B6, AK192, HY6468, PJAK4217, PJAK4298) as described previously^{34 (link)} and were routinely cultured in Roswell Park Memorial Institute (RPMI) 1640 (Invitrogen) 10% FBS (Invitrogen). For inducible Kras derived cell lines, 1 ug/ml of doxycycline was directly added to the media. For metabolic and metabolomic assays, 10% dialysed FBS (Atlanta Biologicals Inc.) was used. The cell lines were mycoplasma free, based on tests done monthly in the laboratory using Lonza’s MycoAlert Mycoplasma Detection Kit assays with confirmatory tests by PCR-based assays.

Human lung cancer cell lines H1299, A549, H2030 and H2023 were obtained from ATCC. These cells were routinely cultured in RPMI 1640 (Life Technologies 11965) supplemented with 10% heat inactivated FBS. The cell lines were authenticated by short tandem repeat (STR) profiling at the Institute for Applied Cancer Sciences, MD Anderson Cancer Center. Murine tumor derived cell lines (KP and KPS) were generated by mechanically mincing lung tumors, dissociation with a combination of collagenase I, IV and dispase. These cells were grown in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% FBS and 1% Pen/Strep (Life Technologies 15140). All cells were routinely tested for mycoplasma by PCR. For metabolic and metabolomic assays, 10% dialysed FBS (Atlanta Biologicals Inc.) was used.