Illumina HiSeq 2000 sequencing; Peripheral positioning of lysosomes supports melanoma aggressiveness
Peripheral positioning of lysosomes supports melanoma aggressiveness
Primary human melanoma cell lines were purchased from Rockland and cultured in MCDB153 (ref. n°M7403, Merck/Sigma-Aldrich) and Leibovitz's L-15 medium (ref. n°11415064, Thermo Fisher Scientific) in a 4 to 1 ratio, supplemented with 2% foetal bovine serum, 1.68 mM CaCl2 and 1% penicillin/streptomycin.  RNA was isolated from melanoma cell lines using the RNeasy Mini Kit (Qiagen, Hilden, Germany). RNA integrity was evaluated on an Agilent Bioanalyzer 2100 using an Agilent RNA 6000 Pico Kit (Agilent Technologies, Santa Clara, CA).  Sequencing libraries were prepared using “NEBNext Ultra II Directional RNA Library Prep Kit for Illumina” combined with “NEB Ultra II polyA mRNA magnetic isolation” for mRNA enrichment (New England Biolabs, Ipswich, MA, USA).