Western Blot Analysis of EMT Markers

The antibodies of EMT-related biomarkers including E-cadherin, N-cadherin, Vimentin, and α-SMA were purchased from Cell Signaling Technology. The antibody of PRKCSH was purchased from Santa Cruz Biotechnology. Briefly, the samples were treated with lysis buffer, subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, and transferred to polyvinylidene fluoride membranes. After the membranes were blocked with 5% nonfat milk in Tris-buffered saline containing Tween-20 for 2 hours, the membranes were incubated with primary and secondary antibodies overnight at 4 °C. An ImageQuant LAS500 system (Molecular Dynamics) was used to visualize the bands. Details of the Western blot analysis can be found in our previous publications.^{28 (link)-31 (link, link, link)}