Bap1 Plays a Survival Role in Cutaneous Melanoma

SUPPLEMENTARY MATERIALS Cell lines and culture conditions The cutaneous melanoma cell lines,(A375, SKmel-28, SKmel-119 and SKmel-63) have been previously reported (Ji et al., 2013; Ji et al., 2012; Tsao et al., 2000) and were maintained in DMEM (Corning life Sciences, Tewksbury, MA) with 10% FBS plus 1x antibiotics. The uveal melanoma cell line, C918, was a kind gift from Dr. Elisabeth Seftor (Children's Memorial Hospital, Chicago, USA) and was maintained in DMEM supplemented with 1X β-mercaptoethanol, antibiotics (Life Technologies, Grand Island, NY), HEPES (Lonza, Walkersville, MD) and 10% FBS (Atlanta Biologicals, Norcross, GA). None of the melanoma lines harbored BAP1 mutations on direct sequencing. The immortalized primary melanocyte line, Pmel (Garraway et al., 2005), was a kind gift of Hans Widlund (Brigham and Women’s Hospital, Boston, MA) and were maintained under the same conditions as the cutaneous melanoma lines. The primary human melanocyte-1 (PHM1) line was purchased from Life Technologies while the primary human melanocyte-2 (PHM2) was a gift of Dr. Mark Pittelkow (Mayo Clinic, Rochester, MN). Both were grown in medium 254 with human melanocyte growth supplement (HMGS) from Life Technologies. MG132 and cycloheximide were purchased from Sigma-Aldrich (Natick, MA). Global gene expression data from primary melanomas came from the study by Harbst et al. (Harbst et al., 2012). BAP1 levels for gene set GDS1375 were downloaded from the Gene Expression Omnibus (GEO; http://www.ncbi.nlm.nih.gov/geo/). Levels of BAP1 for the Cancer Cell Line Encyclopedia (CCLE) were derived from the CCLE website (http://www.broadinstitute.org/ccle/home)