Total RNA was extracted using TRIzol reagent and retrotranscribed after DNase I (Roche, Switzerland) treatment using a FastQuant RT Kit (TIANGEN, P.R. China). Real-time PCR was performed for 45 cycles with SYBR Green PCR Master Mix (TaKaRa, Japan) and processed on CFX 96 system (Bio-Rad, Hercules, CA, USA) as previous described17 (link). Cycle conditions were as follows: initial denaturation at 95°C for 15 min followed by 44 cycles of denaturation at 95°C for 30 s, annealing at 59°C for 30 s, extension at 72°C for 30 s, and plate reading. After the cycles, a dissociation curve was generated from 55°C to 95°C with readings every 0.5°C. Reactions were run in triplicate for each sample. Threshold cycles (Ct) for each tested gene were normalized to the housekeeping β-actin gene value (ΔCt), and every experimental sample was referred to its control (ΔΔCt). Fold change values were expressed as 2−ΔΔCt.