Low glucose

Human cancer cell lines were obtained from ATCC and have not been cultured for longer than 6 months. H1299 (carcinoma, non-small-cell lung cancer) cells were cultured in Roswell Park Memorial Institute (RPMI) medium (Euroclone) supplemented with 10% fetal bovine serum (Euroclone), 1% L-glutamine (Gibco), and 1% penicillin/streptomycin (Euroclone). U-2 OS (osteosarcoma) cells were cultured in low- glucose Dulbecco’s modified Eagle’s (DMEM) medium (Lonza) with 10% fetal bovine serum (Gibco), 1% L-glutamine (Gibco), and 1% penicillin/streptomycin (Gibco). H1299 cells stably overexpressing FLAG-hTERT and hTR were generated through the transfection of the respective linearized vector and were selected using blasticidin (5 mg/mL) and puromycin (1 mg/mL).

PC12 Tet‐Off cells (Clontech, 631134, termed PC12 cells for simplicity) and their derivatives stably expressing Nrg1 were cultured in low glucose DMEM medium (1 g/l, Lonza) supplemented with 10% FCS, 5% HS, 50 μg penicillin, 50 μg streptomycin and GlutaMAX (Life Technologies) at 37°C, and 5% CO₂. T‐47D cells (ATCC:HTB‐133) were grown RPMI‐1640 (Life Technologies) supplemented with 0.2 units/ml bovine insulin (Sigma), 10% FCS, 50 μg penicillin, 50 μg streptomycin and GlutaMAX (all Life Technologies) at 37°C, and 5% CO₂. PC12 cells were grown on poly‐l‐lysine‐coated surfaces for both maintenance and experiments; T‐47D cells were only grown on poly‐l‐lysine‐coated surfaces for experiments. Cell lines were negative for mycoplasma contamination.