Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA110499 | ENA |
REPOSITORIES: ENA
Similar Datasets
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Project description:Both benign nevi and melanomas exhibit high rates of activating mutations in BRAF kinase, suggesting that this event is important for melanocyte proliferation and melanoma initiation in vivo; however, the precise mechanism of BRAF kinase action in these lesions remains to be elucidated. We have used comprehensive gene expression profiling to evaluate the downstream effectors of activated BRAFV600E kinase in primary human melanocytes and identified a dominant proliferative target gene signature for BRAF kinase in these cells. We used microarrays to detail the global gene expression pattern change induced by the activation of RAS/BRAF/MEK/ERK signaling pathway and identified distinct classes of up-regulated genes during this process. Experiment Overall Design: Primary human melanocytes were isolated from neonatal foreskins. Melanocytes isolated from five individual donors were pooled together in order to reduce genetic variations and transduced by BRAFV600E expressing lentivirus and control virus with MOI 10 respectively. Cells were harvested 5 days following infection and total RNA was extracted and purified by RNeasy kit by following manufacturerâs protocol. RNA quality check, double strand complementry DNA synthesis, hybridization with Human Genome U133 Plus 2.0 Array Chips (Affymetrix Inc. Santa Clara, CA), initial data extraction, normalization using RMAExpress software.
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Project description: Not available
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