Genegenius super 12

RT-PCR was used to analyze the level of Nrf2 mRNA. Total RNA was isolated from 5 × 10⁶ treated PC12 cells at logarithmic phase by using RNAiso Plus (Takara, Shiga, Japan) according to the manufacturer’s instructions. Forward and reverse primers were 5′-CCATTTACGGAGACCCAC-3′ and 3′-CTTATTTCAACGGCGAGT-5′ for Nrf2, 5′-AAATGGGTGATGCTGGTG-3′ and 3′-TGAGCGAGTTCTAACAGTCG-5′ for GAPDH [40 (link)]. Reverse transcription used reagents from Promega following the manufacturer’s instructions. The RT-PCR products were separated on 1.2% agarose gel and the intensity of each band was quantified using SynGene software (SynGene, Cambridge, UK) and expressed in arbitrary units (GeneGenius Super 12, Syngene, Cambridge, UK).

RT-PCR was used to analyze the levels of Nrf2, NQO-1, HO-1, CAT, and GAPDH mRNA. GAPDH was used as an internal standard. Total RNA was isolated from 5 × 10⁶ PC12 cells in the logarithmic phase using RNAiso Plus (Takara, Shiga, Japan) according to the manufacturer’s instructions. Reverse transcription was done using ReverTra Ace Master Mix from Toyobo (Osaka, Japan) following the manufacturer’s instructions. Semiquantitative RT-PCR was performed with a thermal cycler system (Bio-Rad) using PrimeSTAR GXL DNA polymerase (Takara, Shiga, Japan). The primers used for PCR are listed in Table 1. The RT-PCR products were separated on 2% agarose gel, and the intensity of each band was quantified using SynGene software (SynGene, Cambridge, UK) and expressed in arbitrary units (GeneGenius Super 12, Syngene, Cambridge, UK).