Schwann cell media scm

Human oral SCC cells (HSC-3) (Japanese Collection of Research Bioresources, passage 4–10) were grown in DMEM (Invitrogen) containing antibiotic (penicillin/streptomycin, 10 U/ml) and 10% fetal bovine serum (FBS). DOK cells (Sigma Aldrich, passage 4–8) were cultured in DMEM with 10% FBS and 5 μg/ml hydrocortisone. Human primary cultures of Schwann cells (ScienCell Research Laboratories) were grown in Schwann Cell Media (SCM, ScienCell Research Laboratories). Cells were authenticated by ATCC and routinely tested for Mycoplasma (PlasmoTest Mycoplasma Detection Kit; InvivoGen).

Rat primary Schwann cells (pSC) were derived from neonate rat sciatic nerves and obtained along with the Schwann cell media (SCM) and Schwann cell growth supplement (SCGS), from ScienCell Research Laboratories™ (Carlsbad, CA). Palmitic acid (PA), DHA, and poly‐L lysine were provided by Sigma‐Aldrich (St. Louis, MO). Fatty acid‐free BSA and LY294002 were purchased from EMD Biosciences (San Diego, CA). MedChem Express provided our NVP‐BKM120 (Princeton, NJ). Annexin V FITC and 7‐amino‐actinomycin D (7AAD) were purchased from BD Biosciences (San Diego, CA) and eBioscience (San Diego, CA) respectively. Finally, we obtained the antibodies to total AKT (mouse, Cat# 2920, http://scicrunch.org/resolver/AB_1147620) and phosphorylated AKT Ser473 (rabbit, Cat# 4060, http://scicrunch.org/resolver/AB_2315049) or Thr308 (rabbit, Cat# 2965, http://scicrunch.org/resolver/AB_2255933), and rapamycin from Cell signaling Technology (Danver, MA), the actin (mouse, Cat# A5441, http://scicrunch.org/resolver/AB_476744) from Sigma‐Aldrich (St Louis, MO) and Torin 1 from Tocris Bioscience (Bristol, UK). All of these antibodies are commercially available see characterization details in Table 1.