Quantitative PCR
Total RNA was isolated using a NucleoSpin RNA kit (Takara Bio Inc.,
Shiga, Japan). RNA was reverse transcribed using a PrimeScript RT
reagent kit with a gDNA eraser (Takara Bio Inc.). cDNA was amplified
with a TB Green Premix Ex Taq II kit (Takara Bio Inc.) using a Thermal
Cycler Dice Real Time System (Takara Bio Inc.). All primers were
purchased from Takara Bio Inc. The primer sequences are listed in Table
S1. Relative mRNA levels were normalized to Gapdh mRNA levels.