将卵巢癌细胞以2000个细胞/孔接种在96孔板中并孵育过夜,然后用0-160μM山奈酚处理24小时,一式三份。除去培养基,将板冷冻解冻以裂解细胞。向每个孔中加入含有5x SYBR Green I的200μL1×CyQUANT细胞裂解缓冲液,并在室温(RT)下孵育5分钟。将反应(50μL)转移至PCR条管,并使用实时Chromo4 PCR仪器在90℃下测量荧光信号。为了确保细胞增殖测定在细胞数的线性范围内进行,通过在96孔板中接种不同量的OVCAR-3细胞(基于用血细胞计数器计数)产生标准曲线,并测量基因组DNA丰度。过夜孵化后。进行三次独立实验,汇总数据进行统计分析[2]。
数据来源文献
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