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doi: 10.1007/s00262-018-2120-5

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Total RNA was extracted from the cells of each treatment group by the TRIzol method, and the cDNA was synthesized by reverse transcription using total RNA as a template with Reverse Transcription Kit (Takara Bio Inc, Japan) at 37°C for 15 min, 85 °C for 5 s, and 4 °C for 10 min