Lentiviral and siRNA transfection

TP Tingzheng Pan
XL Xuetao Li
YL Yanyan Li
ZT Zhennan Tao
HY Hui Yao
YW Yue Wu
GC Guangliang Chen
KZ Kai Zhang
YZ Youxin Zhou
YH Yulun Huang
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GeneChem (China) designed two shRNAs lentivirus against USP7, shUSP7-1: 5’ -UGUAUCUAUUGACUGCCCUTT- 3’ and shUSP7-2: 5’ -UGGAUUUGUGGUUACGUUACUC-3’, and an ARF4 overexpression lentivirus. The constructed lentiviral vector was transfected into cells followed by puromycin intervention, and the transfection efficiency was identified by western blotting analysis to screen for stably transfected cell lines. Sangon Biotech (China) designed two siRNAs against ARF4, siARF4-1: 5’ -GCAAGACAACCAUUCUGUAUATT- 3’ and siARF4-2: 5’ - CCAUCAGUGAAAUGACAGAUATT- 3’. Lipofectamine 3000 was used for co-transfection for 8 h, and transfection efficiency was investigated after transfection.

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