HeLa cell line was obtained from the American Type Culture Collection (ATCC) and grown in DMEM (Thermo-Fisher Scientific) supplemented with 10% FBS (Thermo-Fisher Scientific) and 1% Penicillin/Streptomycin mix (EuroClone). Cells were maintained at 37°C in an atmosphere of 5% CO 2 .
Cells were transfected using polyethylenimine (PEI) (Sigma-Aldrich) in Opti-MEM medium (Thermo-Fisher Scientific), and incubated twice with plasmids and PEI with a 8 h-release between one transfection and the other. After the second transfection cells were incubated in DMEM and treated with 250 μM of the NO donor DETA-NONOate (Enzo Life Sciences) for 22-24 h and/or 200 nM Staurosporine (STS) (Sigma-Aldrich) for 4h.
Cells were transfected using polyethylenimine (PEI) (Sigma-Aldrich) in Opti-MEM medium (Thermo-Fisher Scientific), and incubated twice with plasmids and PEI with a 8 h-release between one transfection and the other. After the second transfection cells were incubated in DMEM and treated with 250 μM of the NO donor DETA-NONOate (Enzo Life Sciences) for 22-24 h and/or 200 nM Staurosporine (STS) (Sigma-Aldrich) for 4h.