L glutamine

MDCK were maintained in DMEM; Cat #10569-010; Gibco. Human PDACs cells: BxPC3, Panc 10.05, and AsPC-1 were maintained in RPMI-1640 (30-2001; ATCC), MiaPaCa2 and PANC-1 were maintained in DMEM, HPAC were maintained in DMEM-F12 (Cat # 11320033; Gibco), and HPAF-II were maintained in EMEM (30-2003; ATTC). Human NSCLCs: H522, H1975, H1299, A549, H23, H441, H1703, and H358 were maintained in RPMI-1640. All cancer cell lines were maintained in media supplemented with 10% FBS (Cat #16000-069; Gibco) and 2 mM L-glutamine (Cat # CA009-010; GenDEPOT). Cells were test for mycoplasma (Cat# LT07-710; MycoAlert PLUS Mycoplasma Detection Kit). All cell lines were maintained in an incubator at 37°C at 5% CO₂.

Madin-Darby canine kidney (MDCK) and baby hamster kidney (BHK) cells were maintained in Dulbecco’s modified eagle medium (DMEM, Cat#10569-010; Gibco). Human non-small cell lung cancer cells (NSCLCs): H1299, H23, A549, H358, and H441 were all maintained in RPMI-1640 (Cat#30-2001; ATCC). Human pancreatic ductal adenocarcinoma cells (PDACs): BxPC3, AsPC-1 and Panc 10.05 were maintained in RPMI-1640, MiaPaCa-2 and PANC-1 were maintained in DMEM and HPAF-II were maintained in EMEM (Cat#20-200; ATCC). All cancer cell lines were maintained in media supplemented with 10% FBS (Cat#16000-044; Gibco) and 2mM L-glutamine (Cat#CA009-010; GenDEPOT). Cells were routinely tested for mycoplasma (Cat#LT07-710; MycoAlert PLUS Mycoplasma Detection Kit; Lonza; Rockland, ME). All cell lines were maintained in 37 °C incubator with 5% CO₂ injection.

T47D cells (ATCC HTB-13) were maintained in RPMI-1640 medium (30–2001, ATCC) supplemented with 10 μg/mL human recombinant insulin (12585–014, Gibco), 2 mM L-glutamine (CA009–010, GenDepot) and 10% FBS (16000044, Life Technologies). Cells were transfected with mGFP-KRASG12V, -HRASG12V or -LactC2 in pEF6 vector (Invitrogen) using TransIT-BrCa Transfection Reagent (MIR 5500, Mirus), and selected with 10 μg/mL blasticidin for 3 weeks. After the selection, cells were maintained in 5 μg/mL blasticidin. For generating MTMR knockdown cell lines, wild-type (WT) T47D cells were infected with lentivirus expressing shRNA targeting MTMR2, 3, 4 or 7. Cells were selected by puromycin (1 μg/mL) for 1 week and maintained in 0.5 μg/mL puromycin. Cells were grown at 37°C in 5% CO₂ and frequently tested for mycoplasma using MycoAlert Mycoplasma Detection Kit (Lonza, LT07–318).