Wm1552c

The SK-Mel147, WM 1366, and SK-Mel173 melanoma cell lines were generously provided by M.S. Soengas (CNIO, Madrid, Spain). The MeWo, SK-Mel131, M17, and TRP melanoma cell lines were used in a previous study [41 (link)]. The WM 1552C, WM 35, WM 793, and WM 115 melanoma cell lines were purchased from ATCC. Cells were cultured in DMEM:F12 medium (1:1) supplemented with 10% fetal bovine serum (Lonza) and maintained at 37 °C in a humidified incubator with 5% CO₂. The analyses were performed within a few passages after thawing. Cells were routinely tested for Mycoplasma contamination.

The human melanoma cell lines WM1552C (Rockland Immunochemicals, Limerick, PA, USA, WM1552C-01-0001), C8161 and UACC1273 (gifts from Dr. Richard Seftor, University of West Virginia), and Sk-Mel28 (ATCC, Manassas, VA, USA, HTB-72) were used. These cells were maintained in RPMI1640 media supplemented with 5% HI-FBS for WM1552C (RPMI: GenClone, San Diego, CA, USA, 25-506H; FBS: Seradigm, Batavia, IL, USA, 97068-085), RPMI1640 media supplemented with 10% HI-FBS for C8161, EMEM (ATCC, 30-2003) with supplemented 10% HI-FBS for Sk-Mel28, and RPMI1640 supplemented with 10% HI-FBS and 0.1% Gentammycin for UACC1273. We also used the primary cell line epidermal keratinocytes (HEKn) (ATCC, PCS-200-010) that were grown in Dermal Cell Basal Media (ATCC, PCS-200-030) and keratinocyte growth supplement (ATCC, PCS-200-040) and the Neuroblastoma cells SH-SY5Y (ATCC, CRL-2266) that were grown in EMEM/F12 containing 10% HI-FBS (EMEM: ATCC 30-2003; F12: Gibco, Waltham, MA, USA, 11765-054). The SH-SY5Y neuroblastoma cells were used as a Netrin-1-expressing positive control, while the HEKn epidermal keratinocytes were used as a poor-Netrin-1-expressing control. Cells were incubated in 37 °C and 5% CO₂ conditions.