Mytaq mix kit

After treated the cells with different concentrations of DEN or DEN-HPβCD, Total RNA was isolated by using a Trizol RNA isolation kit (Invitrogen, Carlsbad, CA, USA) and stored at −80 °C for subsequent analysis. First-strand cDNA was synthesized from 600 ng of RNA by using iScript cDNA Synthesis kit (Bio-Rad). MyTaq Mix kit (Bioline, Taunton, MA, USA) was used for amplification. The sequences of primers used in RT-PCR were shown in Table 1. The PCR cycling conditions for caspase-3 were 95 °C for 1 min and the amplification was followed by 30 cycles of denaturing at 95 °C for 15 s, annealing at 60 °C for 15 s, and primer extension a 72 °C for 10 s with a final extension at 72 °C for 7 min. The PCR cycling conditions for β-actin were 95 °C for 1min and the amplification was followed by 30 cycles of denaturing at 95 °C for 15 s, annealing at 61 °C for 15 s, and primer extension a 72 °C for 10 s with a final extension at 72 °C for 7 min. The PCR product was analysed on 1.5% agarose gels and DNA bands were visualized by 5× loading buffer and Bio rad gel documentation system/Bio-Rad. All experiments were performed in triplicate.