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Reverse transcription–polymerase chain reaction

CW Cenhao Wu
JG Jun Ge
MY Ming Yang
QY Qi Yan
YW Yingjie Wang
HY Hao Yu
HY Huilin Yang
JZ Jun Zou
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Trizol (Invitrogen, Carlsbad, USA) was used to lyse NPCs according to manufacturer's protocol. RevertAid RT Reverse Transcription Kit (TaKaRa Bio, Otsu, Japan) was used to reverse transcribed the total RNA into cDNA as instructed. SYBR Green Master Mix Reagent (Toneker, Shanghai, China) was used for quantitative PCR for determination of mRNA levels (IL-6 and STAT3) according to the protocol. An internal reference gene (GAPDH) was amplified simultaneously with the target genes. Primer sequences were as follows: IL-6-F: 5'-GAGGATACCACTCCCAACAGACC-3', IL-6-R: 5'-AAGTGCATCATCGTTGTTCATACA-3'; STAT3-F: 5'-GAGTCAGGCACTGTGGG-3', STAT3-R: 5'-CGGTCGGTTTCTGCCTGTA-3'; GAPDH-F: 5'-TTCAACGGCACAGTCAAG-3', GAPDH-R: 5'-TGGTTCACACCCATCACA-3'. The experiment was independently repeated three times.

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