Real time PCR was performed using 2× DyNAmo syprogreen qPCR kit (New England Biolabs) according to manufacturers procedures. Briefly, reactions were composed of 5 µl 1.2 µM forward and reverse primers, 5 µl diluted cDNA template and 10 µl 2× qPCR mix (see Table S3 for primer sequences). Reactions were heated for 10 min at 94°C, then cycled for 35 cycles at 20 sec at 94°C, 20 sec at 60°C or 52°C, 30 sec at 72°C. After each cycle the sample was heated to 78°C for 10 sec prior to reading sample fluorescence. Reactions were done in triplicate. ΔΔCt method was used to quantify the relative levels of expression to the Gapdh or β-actin house keeping genes. Expression levels were then normalized to un-induced cells control cells.
