FBXL10 and EMT regulation in breast cancer

The human FBXL10 expression plasmid was used as previously described [43 (link)]. The GFP-SNAI1, HA-SLUG, Flag-ZEB1, Myc-HDAC1 and Myc-HDAC2 were constructed from our lab [58 (link)]. p300 plasmid was mentioned in our previous study [56 (link)]. For HEK293T, polyetherimide was used for transfection reagent. Breast cancer cells were transfected with jetPRIME transfection reagent (Polyplus, 114-15) according to the manufacturer’s specifications. The siRNAs for human FBXL10 were obtained from Hippobiotec, the sequences were listed: 5′-GCAAACAGAGUGACAUCUUTT-3′ (#1 sense), 5′-AAGAUGUCACUCUGUUUGCCT-3′ (#1 antisense); 5′-GGACCCAUCUCACUGAGUUTT-3′ (#2 sense), 5′-AACUCAGUGAGAUGGGUCCAT-3′ (#2 antisense); 5′-CCAUCUGCAAUGAAAUCAUTT-3′ (#3 sense), 5′-AUGAUUUCAUUGCAGAUGGAG-3′ (#3 antisense). The sequences of siRNAs targeted for SNAI1 were listed: 5′-CCACUCAGAUGUCAAGAAGUA-3′ (#1 sense), 5′- UACUUCUUGACAUCUGAGUGG-3′ (#1 antisense); 5′-CCAAUCGGAAGCCUAACUACA-3′ (#2 sense), 5′-UGUAGUUAGGCUUCCGAUUGG-3′ (#2 antisense). The targeted sequence of shRNA for mouse FBXL10 was 5′-GCTGTGGAAATATCTGTCATA-3′, shRNAs were constructed using pRNAT-U6.1/Hygro.