αTC1-6 and βTC1 cells (passages 20-40) were seeded the day before treatment in 60 mm dishes at a density of 4.5 x 10e5 cells. Control cells were grown without any treatment; test cells were treated with a cocktail of inflammatory cytokines containing IL1b 50 IU/ml; IFNg 1000 IU/ml; TNFa 1000 IU/ml
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted with Trizol (Lifetechnologies™, Foster-City, CA, USA), according to manufacturer's instructions.
Label
FAM
Label protocol
PCR assays were performed using TaqMan® Rodent miRNA A+B Cards Set v2.0. Specifically RNA for HT miRNA expression profiling was reverse transcribed into cDNA of 519 mouse-specific and 68 rat-specific miRNAs through Megaplex™ RT Rodent Primer Pool sets, and preamplified through Megaplex™ PreAmp Rodent Primer Pool sets (Lifetechnologies™). Resulting cDNAs were loaded onto TaqMan Low Density miR Arrays (TLDA) cards, according to manufacturer’s instructions.
