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Immunostaining of HBE cells ALI cultures were fixed in 4% paraformaldehyde 20 min at RT, followed by incubation in permeabilization solution (0.3% Triton X-100/PBS) for 15 min and in blocking solution (5% FBS/PBS/3% BSA) for 1 h at RT, incubated in primary antibodies overnight at 4 C, washed, incubated in secondary antibodies for 2 h at RT, washed, and then mounted for imaging