RNA extraction and real-time PCR
Total RNA was prepared using Trizol reagent (Thermo Fisher, USA)
according to the manufacturer’s instructions. RNA purity and
concentration were measured using a spectrophotometer. One microgram of
total RNA was reverse transcribed into cDNA using an
RT Easy II First Strand cDNA
Synthesis Kit (Roche, Cat#04379012001). Then, cDNA(18 ng) was amplified
in a Real-Time PCR Easy (SYBR
Green I) (MCE, Cat#HY-K0501) on Bio-Rad Sequence Detection system
(Bio-Rad, USA). The following primers were listed in the supplementary
table S1.