Fig. 2 Epithelial tight junctions in 3O-C 12 -HSL-treated human cells. (a)
Expression of ZO-3-JAM-A protein complexes in epithelial Caco-2 monolayers.
Cells were treated with 200μM 3O-C 12 -HSL or diluent (0.6% DMSO) as a
control, for 1, 2, 4, or 24 h. Total cell extracts were immunoprecipitated for
JAM-A or ZO-3 and further immunoblotted for ZO-3 and JAM-A, respectively,
using specific antibodies. (b) Visualization of ZO-3 and JAM-A distribution in
epithelial Caco-2 monolayers. Cells were treated with 10 or 200μM 3O-C 12 -HSL
or diluents (0.6% DMSO) as a control, for 4 h. Samples were fixed and stained
with antibodies against ZO-3, JAM-A and Alexa Fluor 594 and 488 secondary
Abs and analyzed by confocal laser scanning microscopy (CLSM). Image size is
67.667.6μm and pixel size is 0.13μm220 Jake Everett et al.