Gene expression analysis (qRT-PCR)

Total RNAs was extracted using the Tri-Reagent (Molecular Research Center) method. Briefly, each sample was homogenized in 1 ml Tri-Reagent solution mixed with 160 μl of chloroform:isoamyl alcohol (24:1). Following centrifugation (10 min, 13.000 rpm, 4°C) total RNAs present in the aqueous phase were precipitated by the addition of 400 μl of isopropanol followed by another centrifugation. Pellets were then washed twice with ethanol 70% and dried prior resuspension in RNAse-free water. For each sample, 1 μg of total RNA treated with DNase was reverse transcribed into cDNA using the RevertAid kit (Thermo scientific). Quantitative PCR analyses were carried out using a LightCycler^® 480 (Roche) and the LC480-SYBR-Green master I reaction mix (Roche). PP2AA3 (PROTEIN PHOSPHATASE 2A SUBUNIT A3) was used as reference gene [62]. Expression levels were calculated using the comparative threshold cycle method. All the primers used are described S2 Table.