Glutamax

hTert-immortalized human fibroblasts (hTert-Fibroblasts) were obtained from Silvia Soddu (Regina Elena Cancer Institute, Italy)^{63 (link)}. As described previously^{14 (link)}, hTert-Fibroblasts were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco), supplemented with heat inactivated 10% FBS (Australian, Gibco), penicillin-streptomycin (Gibco) and GlutaMAX (Gibco), in a 5% CO2 humidified atmosphere, at 37 °C. Human fibroblasts from SMA type I patient (GM00232) and healthy control (GM08333) were obtained from Coriell Institute for Medical Research (Camden, NJ, USA), and cultured in DMEM medium supplemented with 10% FBS, penicillin/streptomycin, and GlutaMAX, in 5% CO2 humidified atmosphere, at 37 °C. For knockdown experiments, cells were transfected with Lipofectamine 2000 (Thermo Fisher Scientific) and a combination of three siRNA-27 duplexes targeting the human SMN1 gene (OriGene), following manufacturer’s instructions. Universal scrambled siRNA duplex was used as negative control. Cells were harvested after 48 h or 72 h post transfection.

hTert-immortalized human fibroblasts, were grown in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Grand Island, NY, USA), supplemented with heat-inactivated 10% FBS (Australian) (Gibco), penicillin-streptomycin (Gibco) and GlutaMAX (Gibco). Human fibroblasts from SMA type I patient (GM00232) and healthy control (GM08333) were obtained from Coriell Institute for Medical Research (Camden, NJ, USA), and grown in DMEM medium supplemented with 10% FBS, penicillin/streptomycin, and GlutaMAX. All cell cultures were maintained at 37 °C in a humidified atmosphere of 5% CO₂. For knockdown experiments, cells were transfected with Lipofectamine 2000 (Thermo Fisher Scientific) and a combination of three siRNA-27 duplexes targeting the human SMN1 gene (OriGene, Rockville, MD, USA), following manufacturer’s instructions. Universal scrambled siRNA duplex was used as negative control. Cells were harvested after 48 h post transfection.