The total RNA from cardiac rat tissue was isolated with RNA-Bee (Amsbio, UK) and a quantitative real-time PCR was performed with Brilliant SYBR Green qPCR master mix (Applied Biosystems, USA). The relative amount of target mRNA was determined using the comparative threshold (Ct) method as previously described (53 (link)). The mRNA contents of target genes were normalized to the expression of hypoxanthine phosphoribosyl transferase (HPRT).
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