Isolation of intestine epithelial cells from mice
Intestine epithelial cells were prepared as previously
described23. Briefly, precooled PBS (containing 5%
penicillin, and streptomycin) was used to wash the colons of each group
of mice, and ophthalmic shears were used to remove the Pyle’s node, fat,
and mesentery on the surface of the small intestine, and the tissue was
cut to the size of 1cm. Incubate with 1 mM DTT and 3 mM EDTA at room
temperature for 1h without shaking. Cells were washed with PBS by
centrifuging and then were solubilized in cell lysis buffer containing
1% Triton X-100, 1mM EDTA, 1 mM EGTA, 10 mM Tris (pH 7.4), 150 mM NaCl,
and protease, and phosphatase inhibitor cocktail (Solarbio).