For analysis, total RNA was isolated from the entire above organs and cells using the TRIzol RNA Extraction Kit (Invitrogen, CA, USA), according to the manufacturer’s instructions, and the RNA pellets were dissolved in 10 ml of RNase-free distilled water. Total RNA was measured at 260/280 nm using a UV spectrophotometer and was stored at −80 °C prior to use. cDNAs were prepared from 10 μg of total RNA in a 100 μl reaction mixture using random primers according to a standard protocol (high capacity cDNA archive kit; PE Applied Biosystems, Foster City, CA, USA). The PCR reactions were conducted using an iCycler thermal cycler (Bio-Rad, Hercules, CA, USA), and the mixtures were stored at −80 °C until analysis. Real-time RT-PCR was performed using 3 ng of cDNA and the validated SYBR Green gene expression assay in combination with the SYBR Premix Ex Taq II (TaKaRa, Bio Inc., Ohtsu, Japan) for measuring Tubb2c, Atp6v1a, Pdhb, Hsc70t, Fbp1, Lrrc34, Tyrp1, Gapdhs, Pdha2, Dazap1, Fh1, and GAPDH. All of the primers used in this analysis are listed in Table 4. Quantitative real-time PCR was performed in duplicate in a thermal cycler dice real time system TP800 (TaKaRa). The data were analyzed using thermal cycler dice real time system software (TaKaRa), and the comparative Ct method (2∆∆Ct) was used to quantify gene expression levels. Real-time RT-PCR data were standardized to GAPDH, which was used as an internal control. To confirm the specific amplification of the target genes, each gene product was further separated on a 1.5% agarose gel to detect any single bands at the theoretical product sizes, and the dissociation curves were analyzed to detect any single peaks.