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Total RNA was extracted using TRIZOL (Invitrogen). The cDNA was reverse transcribed from total RNA using the Prime Script RT reagent kit (Takara Bio, Otsu, Japan). Quantitative RT–PCR was performed using SYBR Green I (Takara Bio) and the Light Cycler real-time PCR apparatus (Roche). Primer sequences are listed in Supplementary Table 1, statistical analysis was performed using the two-tailed Student’s t-test or analysis of variance.Chikuma Erotic Massage
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