Mcdb153 medium

Unless otherwise specified, reagents were obtained from MilliporeSigma (Burlington, MA, USA). The melanoma cell lines WM35, WM164, WM793 were provided by M. Herlyn (Wistar Inst., Philadelphia, PA, USA). These cell lines and variants derived from them were cultured in MCDB153 medium supplemented with 2% FBS (Invitrogen, Carlsbad, CA, USA), 2.5 µg/mL insulin solution and 2 mM CaCl₂ (Tu2%). The WM35 line was originally derived from a melanoma in radial growth phase (RGP), WM793 was isolated from a vertical growth phase (VGP) melanoma, and WM164 was derived from a metastatic lesion. WM793 clones expressing FLAG-NME1 or FLAG only (“empty vector” or FLAG only) were previously described [8 (link),31 (link)]. The head and neck squamous cell carcinoma (HSSCC) cell line Tu167 was obtained from the University of Texas MD Anderson Cancer Center (Houston, TX, USA), and was cultured in complete Dulbecco’s Modified Eagle Medium (Invitrogen, high glucose) supplemented with 10% FBS (R&D Systems, Minneapolis, MN, USA), 2 mM Glutamax (Gibco, Carlsbad, CA, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL).

Normal human melanocyte cell line NHEM was cultured in Ham's F10 media supplemented with ITS premix (Becton Dickinson, Franklin Lakes, NJ). Human melanoma cell lines WM793B, WM35, A2058, A375 and 451Lu were obtained from American Type Culture Collection (ATCC, Manassas, VA). WM35 and 451Lu cell lines were cultured in RPMI 1640 medium (Invitrogen) supplemented with 10% foetal bovine serum (Invitrogen). WM793B cell line was maintained in MCDB153 medium with 10% foetal bovine serum (Invitrogen). A2058 and A375 cell lines were cultured in Dulbecco's MEMV F12 medium (Invitrogen) supplemented with 10% foetal bovine serum (Invitrogen). Cisplatin (Sigma‐Aldrich, St Louis, MO) was used at a concentration of 25 μmol/L unless specified description.