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.