Cellular Immunofluorescence Microscopy Protocol

Cellular immunofluorescence was performed as we described previously^{17 (link)}. Cells were seeded into the 6-well culture plate (Corning Costar Corp) to prepare for performing cell immunofluorescence (IF). After incubating with primary antibodies, cells then incubated with corresponding fluorescence labeled secondary antibody. After stained with DAPI (Beyotime Institue of Biotecchnology, Jiangsu, China), the slides were photographed using the inverted fluorescence microscope TE-2000S (Nikon, Tokyo, Japan). Primary antibodies for E-cadherin, vimentin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). All fluorescence labeled secondary antibody were obtained from Beyotime Institute of Biotechnology (Shanghai, China). Rhodamine-conjugated phalloidin (Beyotime Institute of Biotechnology) was used to analyze to cytoskeleton. Cells grown on cover slides were fixed, then incubated with Rhodamine-conjugated phalloidin (Beyotime Institue of Biotecchnology) and followed by DAPI (Beyotime Institue of Biotecchnology).