Total protein was extracted with RIPA buffer (Solarbio). After the protein was quantified using BCA™ Protein Assay Kit (Pierce, Appleton, WI, USA), the equal amounts of protein samples were separated by polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% fat-free milk for 2 h and incubated with primary antibodies against E-cadherin (1:500, ab15148, Abcam), Vimentin (1:2,000, ab137321, Abcam), CBL (1:5,000, ab32027, Abcam), or β-actin (1:2,000, ab8227, Abcam). After washing thrice with tris-buffered saline, the membranes were probed with horseradish peroxidase-labeled secondary antibody (1:25,000, ab205718, Abcam). Finally, the protein bands were measured using the ECL system (Beyotime, Shanghai, China).

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