Gm08333

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.

Human dermal fibroblasts (GM08333) were purchased from the Coriell Institute for Medical Research (Camden, NJ, USA). The fibroblasts were cultured in Dulbecco’s Modified Eagle’s medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA, USA). For the 3-D reconstruction of human skin tissue, human neonatal dermal fibroblast (HDFn) was cultured in Medium 106 (M106) supplemented with low-serum growth supplement (LSGS). Human neonatal epidermal keratinocyte (HEKn) was cultured in EpiLife medium supplemented with human keratinocyte growth supplement (HKGS). Penicillin (100 U/mL) and streptomycin (100 mg/mL) were added to the medium as antibiotics, and the cells were cultured at 37 °C in an atmosphere containing 5% CO₂. All cells, media, supplements, and antibiotics were purchased from Thermo Fisher Scientific (Waltham, MA, USA).