Fig 3.

(A) Polarized T-84 cells were treated with either flagellin (0.01 μg/ml) or PBS in the basolateral chamber. Simultaneously, curli fibrils (10 μg/ml) or the synthetic TLR2 ligand Pam₃CSK₄ (0.1 μg/ml) was added to the apical chamber. (B) Polarized T-84 cells were pretreated with the specific PI3K inhibitor LY294002 (10 μM) for 1 h prior to the experiment. Cells were then treated with flagellin (0.01 μg/ml) in the basolateral chamber alone, or simultaneously curli fibrils (10 μg/ml) or the synthetic TLR2 ligand Pam₃CSK₄ (0.1 μg/ml) was added to the apical chamber. Epithelial permeability was determined by treatment for 2 h with FITC-dextran added after 24 h to the apical chamber. Fluorescence in the basolateral supernatants using a BMG Omega plate reader was determined.