Abi q6 apparatus

Total RNA was extracted from the spleen and brain with RNAiso plus (TaKaRa Biotechnology, Beijing, China). The mRNA was reverse transcribed into cDNA using a PrimeScript RT reagent kit (TaKaRa Biotechnology, Beijing, China). Quantitative real-time PCR was performed by using appropriate primers mentioned in Supplemental Table 1 and SYBR green PCR master mix (Roche, Penzberg, Germany) for 40 cycles each in an ABI Q6 apparatus (Applied Biosystems, CA, USA). The mRNAs expressions were calculated using the ratio of the gene of interest to β-actin. All qRT-PCR assays were performed in triplicate.

Total RNA was extracted from the hearts with RNAiso plus (TaKaRa Biotechnology, Beijing, China). The mRNA was reverse transcribed into cDNA using a RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific, USA). Quantitative real-time PCR was performed using SYBR green PCR master mix (Roche, Penzberg, Germany) for 40 cycles in an ABI Q6 apparatus (Applied Biosystems, CA, USA). Real-time fluorescence quantitative PCR was performed using the following primers: Gapdh forwards primer, 5′-TGGCACCACACCTTCTACAATG-3′; Gapdh reverse primer, 5′-TCATCTTCTCGCGGTTGGC-3′; Scn5a forwards primer, 5′-GAAGTGCCAGCGTAACTCCTA-3′; and Scn5a reverse primer, 5′-TACCTCAAACACCCCATTCAC-3′.