Regulation of SNAI2 by USP7 and TRIM21

All transfection experiments were performed as previously described [47 (link)]. Plasmids shSNAI2, shUSP7 and shTRIM21 and their lentiviral packaging were commissioned for Genepharma (Shanghai, China) synthesis, and overexpressed plasmids (FLAG-tagged SNAI2, FLAG- or HA- or Myc-tagged USP7/wt, Myc-tagged USP7/C223S, HA-Ub/mt, HA-Ub/wt, Myc-tagged TRIM21) and their lentiviral packaging were commissioned for OBiO (Shanghai, China). siUSP7 and negative controls were purchased from GenePharma. RD and A204 cells were infected with leniviruses, and stable cells were screened with 2ug puromycin. The shRNA sequences used were as follows: shNC:5’-GTTCTCCGAACGTGTCACGT-3’;shSNAI2:5’-CCAGAATGTCTCTCCTGC-3’;shUSP7:5’-CCTGGATTTGTGGTTACGTTA-3’;shTRIM21:5’-TGAGAAGTTGGAAGTGGAAAT-3’; siNC: 5’-UUCUCCGAACGUGUCACGU-3’; USP7 siRNA-1:5’-ACCCUUGGACAAUAUUCCUTT-3’;USP7 siRNA-2: 5’-AGUCGUUCAGUCGUCGUAU-3’.