2.9 | Immunofluorescent staining
The HNECs were seeded in 6-well plates to perform immunofluorescent
staining of the TJ of CRSwNP, CRSsNP, and control. When the degree of
cell fusion was 100%, the cells were fixed with 4% paraformaldehyde
for 15 min, and incubated overnight with ZO-1 antibody at 4 °C. On the
next day, the specimens were washed with PBS, and incubated with
secondary antibodies in the dark at room temperature for 1 h. After
rinsing with PBS, the specimens were incubated with DAPI at room
temperature for 10 min. The slices were sealed using nail polish, and
the specimens were visualised under a fluorescence microscope (Olympus,
Japan).