Anti crabp2

Cells were seeded into 6-cm dishes at a density of 2×10⁵ cells/dish. The cells were fixed with 4% paraformaldehyde for 20 min at room temperature, permeabilized in 0.5% Triton X-100 for 1 h and blocked with 3% BSA at room temperature for 2 h (Nanjing KeyGen Biotech Co., Ltd.). Subsequently, the cells were incubated with the following primary antibodies for 6 h at 4°C in humid conditions: Anti-E-cadherin (1:100; product no. 14472S; Cell Signaling Technology, Inc.), anti-vimentin (1:100; product no. 5741S; Cell Signaling Technology, Inc.) and anti-CRABP2 (1:250; product code ab211927; Abcam). Following the primary antibody incubation, the cells were incubated with Alexa Fluor-594 anti-rabbit (1:500; cat. no. A-11012; Thermo Fisher Scientific, Inc.) or anti-mouse secondary antibodies (1:500; cat. no. A-11032; Thermo Fisher Scientific, Inc.) at room temperature for 1 h. The cell nuclei were subsequently stained with 5 µg/ml DAPI (Beyotime Institute for Biotechnology) for 5 min. Stained cells were visualized using an immunofluorescence microscope at a magnification of ×400 (Olympus Corporation).