Immunofluorescence Staining of Tight Junctions

Paraffin-embedded sections were dewaxed, hydrated, treated for antigen retrieval, and blocked with 10% donkey serum. Then, the primary antibodies were used for the incubation of the sections at 4 °C overnight. The next day, the slides were stained with relevant secondary antibodies at room temperature for 1 h. Nuclei were stained with DAPI (Servicebio, Wuhan, China). Finally, the sections were observed with a confocal microscope (Olympus, Tokyo, Japan) after sealing with an anti-fluorescence quencher. The antibodies used in this study were as follows: anti-ZO-1 (1:200, Genetex, Texas, America), anti-Occludin (1:200, Genetex), and Alexa Fluor 488-conjugated donkey anti-rabbit IgG (1:200, Antgene, Wuhan, China).