WM3704 Purified RNA

Total RNA was extracted and purified from cells using RNA isolation & purification kit according to manufacturers instructions. The RNA was diluted to 200 ng/µL in RNase free water. Concentration was determined at A260 using nanodrop ND-1000. Total RNA was prepared from cell line WM3704. WM3704 is a metastatic human melanoma cell line that displays a mesenchymal morphology. This cell line was established from a lymph node metastasis site of a patient. This cell line features the specific V600E (Val600Glu) mutation at codon 600 in the BRAF gene. This mutation causes constitutively active kinase activity and activation of MEK and ERK signaling pathway. This cell line also expresses mutated PTEN gene and R24H mutation at position 24 in the CDK4 gene. The R24H mutation results in an amino acid substitution at codon 24 in CDK4, from an arginine (R) to histidine (H). This mutation is an activating mutation, as it results in growth advantages by preventing p16 binding, while maintaining the ability of CDK4 to form a functional kinase with cyclin D. WM3704 cells produce xenograft tumors when injected into immunocompromised mice.