RNA Isolation and Real-time Quantitative PCR
Total RNAs from tissues or cells were extracted with Trizol reagent
(Thermo Fisher Scientific Inc, MA, USA). Reverse transcription of 2 μg
total RNA was performed with a high-capacity cDNA reverse transcription
kit (Promega, Madison, USA). qPCR reaction was performed with a SYBR
Green Master Mix (Promega, Madison, USA) and detected using a Prism
VIIA7 real-time PCR system (Applied Biosystems Inc, Foster City, CA,
USA). Primers were designed using Primer Quest (Integrated DNA
Technologies, Inc, Coralville, IA, USA). See supplementary table 10 for
the details of primer sequences.