Lentiviral infection and siRNA transfection were used to knockdown
ZNF3. For lentiviral infection, three ZNF3-shRNA plasmids and control plasmid were constructed and verified by sequencing and then transfected with HEK-293T cells for packaging of the lentivirus virus. Lentiviral particles were collected to infect SW480 cells, followed by puromycin selection to obtain stable
ZNF3-knockdown cell lines. The sequences of ZNF3 shRNAs and siRNAs were as follows: siZNF3#1: 5′-GGAACCTCAGGCCCTGCTTGA-3′, siZNF3#2: 5′-GGAAGGAGTGGAAGCGTTTGG-3′, siZNF3 #3: 5′-GGTAATGCTGGCATGGCTTAC-3′, and control: 5′-TTCTCCGAACGTGTCACGT-3′. ZNF3 overexpression constructs were obtained by using the pEX-3 (pGCMV/MCS/Neo) vector (GenePharma, Shanghai, China), TWIST and MMP1 overexpression constructs were obtained by using the pcDNA3.1 vector (GenePharma), and an empty vector (EV) was used as the control. The Myc flag was provided by Dr. Guo Li at the Department of Biochemistry and Molecular Biology of Hainan Medical University (Haikou, China). Transfection was carried out using Lipofectamine 3000 Transfection Reagent (Invitrogen, Carlsbad, USA).
ZNF3. For lentiviral infection, three ZNF3-shRNA plasmids and control plasmid were constructed and verified by sequencing and then transfected with HEK-293T cells for packaging of the lentivirus virus. Lentiviral particles were collected to infect SW480 cells, followed by puromycin selection to obtain stable
ZNF3-knockdown cell lines. The sequences of ZNF3 shRNAs and siRNAs were as follows: siZNF3#1: 5′-GGAACCTCAGGCCCTGCTTGA-3′, siZNF3#2: 5′-GGAAGGAGTGGAAGCGTTTGG-3′, siZNF3 #3: 5′-GGTAATGCTGGCATGGCTTAC-3′, and control: 5′-TTCTCCGAACGTGTCACGT-3′. ZNF3 overexpression constructs were obtained by using the pEX-3 (pGCMV/MCS/Neo) vector (GenePharma, Shanghai, China), TWIST and MMP1 overexpression constructs were obtained by using the pcDNA3.1 vector (GenePharma), and an empty vector (EV) was used as the control. The Myc flag was provided by Dr. Guo Li at the Department of Biochemistry and Molecular Biology of Hainan Medical University (Haikou, China). Transfection was carried out using Lipofectamine 3000 Transfection Reagent (Invitrogen, Carlsbad, USA).