Semi-quantitative RT-PCR was conducted with purified RNA from mammalian cells [30 (link), 31 (link)]. One μg of RNAs purified using RNeasy kit (Qiagen) was used for synthesis of the first strand cDNA with a random hexamer (Roche) and Superscript III first-strand synthesis kit (Invitrogen). PCR was conducted with high fidelity 2X PCR master mix (NEB).
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