Global microRNA profiling of aTC1-6 and bTC1 before and after treatment with cytokines

The study sought to determine the global miRNA profiles of mouse pancreatic cell lines aTC1-6 and bTC1 at steady state and after treatment with a cocktail of pro-inflammatory cytokines (IL1b; IFNg and TNFa) for a time course of 24 and 48 hours. Inflammatory cocktail contains IL1b 50 IU/ml; IFNg 1000 IU/ml; TNFa 1000 IU/ml. MicroRNA expression profiles in both cell lines during the different experimental conditions were determined using TaqMan® Rodent miRNA A+B Cards Set v2.0 (TLDA, Life Technologies). MicroRNA profiles were measured in 3 independent biological replicates. qRT-PCR analysis of microRNAs of aTC1-6 treated with cytokines for 24 and 48 h and of their respective not-treated controls as well as of βTC1 cells not treated with cytokines. 180 ng RNA of each sample was reverse transcribed through Megaplex™ RT Rodent Primer Pool sets A and B, and preamplified through Megaplex™ PreAmp Rodent Primer Pool sets (Lifetechnologies™). Complementary DNA (cDNA) was amplified usingTaqMan® Rodent miRNA A+B Cards Set v2.0 (Life Technologies™) with TaqMan Universal PCR Master Mix on an ABI 7900HT Sequence Detection System.