Authors
Janoueix-Lerosey Isabelle, Lequin Delphine, and Delattre Olivier
Summary
Genome-wide comparative genomic hybridization analysis and subsequent direct sequencing of neuroblastoma cell lines and primary tumour DNAs allowed us to identify somatic mutations of the ALK-kinase domain. Germline mutations were observed in two neuroblastoma families indicating that ALK is a neuroblastoma predisposition gene. Mutated ALK proteins were over-expressed, hyperphosphorylated and exhibited constitutive kinase activity. The knock-down of ALK expression in ALK-mutated cells, but also in cell lines over-expressing a wild-type ALK, led to a dramatic decrease of cell proliferation. Here, we provide information about the reagents used in this study and describe the RNA interference method, immunoprecipitation protocol and in vitro kinase assay.Further details
The protocol was published on Protocol Exchange in 2008. To see the entire protocol, click on the source link.Advertisement
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