Kifc1

Ovarian cancer cells were lysed using RIPA lysis buffer (Beyotime, China) with the addition of a protease inhibitor cocktail (Thermo Fisher Scientific), and total protein concentration was detected using a BCA Protein Assay kit (Pierce, Rockland, IL, USA). Extracted proteins were separated by 12% SDS-PAGE gel electrophoresis and then transferred onto polyvinylidene difluoride membranes (Roche, Basel, Switzerland). After blocking with dried skimmed milk powder dissolved in TBST solution, the membranes were incubated with specific primary antibodies against KIFC1, CENPE, E-cadherin, N-cadherin, Snail, ZEB1, matrix metalloproteinase (MMP)2, and MMP9 (Cell Signaling Technology) at 4°C overnight. Subsequently, membranes were cultured with peroxidase-conjugated secondary antibodies (Cell Signaling Technology) at room temperature for 2 h. Chemiluminescence (Millipore Corporation) and densitometry analysis by ImageJ software were applied to measure the protein expression.