shRNA and Lentivirus Production

The shRNA sequences were listed in Supplementary Table S2. shRNAs were cloned into SFLV-shRNA-EGFP vector using miR30 primers [23]. HEK 293 T cells were cultured in DMEM medium (Dulbecco’s Modified Eagle Medium) supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/ml), and streptomycin (100 μg/ml). Lentivirus were generated in HEK 293 T cells using calcium phosphate transfection of 20 μg shRNA plasmid, 15 μg pCMVΔR8.91 helper plasmid and 6 μg pMD.G plasmid according to standard procedures [23, 24]. Culture medium was changed 12 h after transfection and virus supernatant was collected 36 h after changing medium. Virus was then concentrated by centrifugation at 25,000 rpm for 2.5 h, 4 °C, and viral pellets were responded in sterile PBS.