All the reactions were performed in duplicate, with the results normalized using the comparative 2−DΔCT method (Livak and Schmittgen, 2001) with the Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) used as housekeeping gene. The data were analyzed using StepOnePlus™ qPCR Systems (Applied Biosystems, Carlsbad, CA, USA).