Lentivirus plasmid construction

The negative control and lentivirus containing the GSG2 (GSG2-Flag) overexpression and knockdown sequences (5’-CCACAGGACAATGCTGAACTT-3’ for GSG2; 5’-GCTGAAGTGAAGAGGCTCAAA-3’, 5’-AGGCAGCTAGAATTGGAATCA-3’, 5’-AAGCTCAGAAAGAGCCATGTT-3’ for KIF15) were purchased from Shanghai Yibeirui Biomedical Science and Technology Co., Ltd. (Shanghai, China). The overexpression sequences and shRNAs were subsequently cloned into pGCSIL-green fluorescent protein lentivirual vector BR-V-108 to generate recombined lentiviral vectors. Lentiviral vectors and packaging vectors were transfected into 293T cells using Lipofectamine 2000 (Invitrogen, New York, CA, USA) according to manufacturer’s instructions. Lentiviral particles were purified using ultracentrifugation, and an endpoint dilution assay was performed to determine the titer of the lentiviruses.