the effect of NT5E enhancing miRNAs was studied by gene expression profiling to unterstand mechanism of observed NT5E upregulation.
Overall design
Gene expression profiling of tumor cell lines transfected with miRNAs enhancing NT5E expression was performed as follows: SK-Mel-28 cells were transfected with 50 nM miRNA, including a non-targeting control miRNA (control-1/ath-miR-416) in a 12 well plate format, performing triplicates for each format. After 48 h, cells were harvested for RNA isolation using an RNAeasy kit (Qiagen, Hilden, Germany). Microarrays were performed by DKFZ-Genomics and Proteomics Core Facility using an Affymetrix Clariom S human chip for all samples. Raw data processing was also performed using the Core Facility. Differential gene expression was calculated by comparing miRNA-induced gene expression levels to the respective gene expression levels following mimic control-1 transfection.
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