Gene array experiment to identify genes that are upregulated by saturating creatine levels in vitro and potentially inhibiting creatine transporter (CrT) activity

We sought to investigate the mechanisms of CrT modulation by saturating extracellular Cr. For this purpose, we used an in vitro system of 3T3 fibroblasts stably overexpressing CrT and followed a global gene array approach to test the hypothesis that saturating intracellular [Cr] increases the expression of candidate molecules that inhibit CrT activity. In this way, we would elucidate mechanisms of redox regulation in cases of fluctuating energy demands. 3T3-CrT cells were grown on 6-well plates to 70% confluency. n=3 replicate wells were treated with 5mM creatine monohydrate in growth media and n=3 wells remained untreated (controls).