A14225

The proteins were abstracted with RIPA (R0010, Solarbio), separated by SDS-polyacrylamide gel system, and electrically transferred to PVDF membranes (IPVH00010, Millipore, USA). After blocking with 5% fat-free milk (A600669, Sangon) for 1 h, the proteins were incubated at 4 °C for 18 hours with the following antibodies: SGK1 (1: 400, A1025, ABclonal, China), cyclin D (1: 1000, A19038, ABclonal, China), cyclin E (1: 500, A14225, ABclonal), N-cadherin (1 : 1000, A19083, ABclonal), Vimentin (1 : 1000, AF7013, Affinity, China), caspase-3 (1 : 1000, CST, No. 14220), caspase-9 (1 : 1000, CST, No. 9508), MMP2 (1 : 500, 10373-2-AP, Proteintech), MMP9 (1 : 1000, 10375-2-AP, Proteintech), E-cadherin (1 : 500, A3044, ABclonal), p-IκBα (1 : 500, AP0707, ABclonal), IκBα (1 : 500, A1187, ABclonal), p65 (1 : 1000, A2547, ABclonal), and p-p65 (1 : 1000, AF2006, Affinity). The next day, the proteins were immuno-blotted with HRP-IgG at 37 ℃ for 1 h. The relative protein level was normalized to GAPDH (1 : 10000, sc-47778, Santa Cruz). Bands were visualized by a gel image system (WD-9413B, LIU YI, Beijing, China).