Hematoxylin-and-eosin (HE) and immunofluorescence
(IF) staining of IECs
Cells (passage 8) were fixed with ice-cold acetone for 10 min,
then hematoxylin-and-eosin (HE) and immunofluorescence (IF)
staining were performed. The major steps of IF staining were
shown as follows. The cells were incubated with rabbit polyclonal
anti-cytokeratin 18 (1:100; Santa Cruz) at 37uC for 1 h, followed
by washing with phosphate-buffered saline (PBS). The cells were
then incubated with FITC-conjugated goat anti-rabbit IgG (1:100;
Santa Cruz), and nuclei were stained with propidium iodide (PI).
In negative control groups, anti-cytokeratin 18 antibodies were
replaced with PBS.