Total rna isolation kit

qRT-PCRs were conducted as previously described [18 (link)]. Total RNA was extracted using a total RNA isolation kit (GeneALL Biotechnology, Seoul, South Korea) according to the manufacturer’s instructions. Then, total RNA was quantified using a NanoDrop spectrophotometer. cDNA was synthesized using a PrimeScript 1st Strand cDNA Synthesis Kit (TaKaRa, Osaka, Japan). qRT-PCR analysis was performed using SYBR Green Master Mix (Roche, Switzerland) and a Light Cycler 480 Real-Time PCR System (Roche, Switzerland). The PCR conditions were set as follows: 94 °C for 1 min (denaturation), 55–58 °C for 30 s (annealing), and 72 °C for 1 min (extension). All primers used in this study are listed in Table 1. The resulting products were normalized using β-actin.

Total RNAs were extracted from cells using a Total RNA Isolation Kit (GeneALL Biotechnology, Seoul, South Korea) according to the manufacturer’s instruction. cDNAs were synthesized using the PrimeScript 1st strand cDNA synthesis Kit (TaKaRa, Osaka, Japan). Quantitative real-time PCR was performed using SYBR Green working solution iTaqTM Universal SYBR Green Supermix (Bio-Rad, Hercules, CA) using a custom PCR master mix and the following conditions: 40 cycles of 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 30 s. mRNA expression levels were normalized to Gapdh expression. The primer sequences were listed in Supplementary Table 1.