Ecm0728l

H1299 human non-small cell lung cancer, HT1080 human fibrosarcoma, HCT116 human colon, and HeLa human epithelial cervix carcinoma cell lines were purchased from American Type Culture Collection. HeLa and HCT116 were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Euroclone, ECM0728L), H1299 and HT1080 in Roswell Park Memorial Institute medium (RPMI 1640; Gibco, 21875–034), both supplemented with 10% fetal bovine serum. The indicated cell lines were grown in a CO₂ humidified incubator at 37 °C. HT1080 and H1299 cells were stably transfected with EGFP-LC3B or mRFP-EGFP-LC3B fusion proteins as previously reported^{38 (link),39 (link)} and cultured in the presence of 800 μg/ml of geneticin (G418 disulfate salt, Sigma-Aldrich, A1720).

Normal mouse embryonic fibroblasts (NHI 3T3) and human lung carcinoma cells (A549) were obtained by American Type Culture Collection (Manassas, VA, USA). The employed cell lines were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) (ECM0728L, Euroclone, Pero, Italy) containing 10% FBS (ECS0180L, Euroclone) and 100 mg/mL penicillin-streptomycin (ECB3001D, Euroclone), and kept in a controlled temperature and atmosphere (37 °C and 5% CO₂).

MDA-MB-231 and MDA-MB-468 human triple negative breast cancer (TNBC) cell lines were purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA) and were cultured in Dulbecco's modified Eagle's medium (DMEM; EuroClone, Italy, ECM0728L), supplemented with L-glutamine, Penicillin/Streptomycin and 10% fetal bovine serum (FBS; Thermo Fisher Scientific - Gibco, Waltham, Massachusetts, USA, 10270-106), in a CO₂-humidified incubator at 37°C.
Stable TRF2 over-expressing and TRF2 silenced cells, and their respective control counterparts (pBabe and shSCR), were obtained as previously described [18 (link)]. For the transient expression of EGFP-LC3B, the cells were transfected for 24 hours by using JetPEI (Polyplus, Illkirch, France, 101-10N), according to the manufacturer's instructions. MDA-MB-231 LUC cells were obtained with a stable infection with lentiviral luciferase vector pRRLSIN.cPPTLuciferase (Addgene, Watertown, Massachusetts, USA).