Total RNA Extraction and Quantification

Total RNA was extracted from mice tissues or primary cultural cells using the RNeasy Lipid Tissue Mini Kit (Qiagen, Germantown, MD, USA). The purity and concentration of total RNA were determined by a NanoDrop spectrophotometer (Thermo Fisher Scientific). One μg of total RNA was reverse-transcribed using a cDNA kit (AB Applied Biosystems, Waltham, MA, USA). Real-time PCR amplification was detected using SYBR Green PCR master mixture (Qiagen) on a Roche 480 Real-time PCR system (Basel, Switzerland). The primer sequences used in real-time PCR are shown in Supplementary Table S1. Western blot analysis was performed following the procedures described previously [45 (link)].