Lentivector information

The lentivector Psico was purchased from Addgene (#11578, Addgene, MA, USA). After being double digested with HpaI /XhoI, the vector was connected with synthetic SP1 knockdown or NC sequences. After screened through sequencing, the positive lentivectors carrying SP1 knock-down sequence or NC were selected for cell infection. PCI-37B cells cultivated in 6-well plate at a confluence of 70% were cocultured with lentivirus suspension (SP1 knock-down 1:100; NC 1:200) for 48 hours. Then cells were cultured with complete culture medium with 15ug/ml puromycin for one week to harvest strains that have stably low levels of SP1 expression.