JAM1 is required for epithelial barrier function of IHGE cells.
(A) Schematic image of the culture insert system. Monolayer of IHGE cells stably expressing shLuc, shJAM1 #110, or shJAM1 #508 were cultured in culture inserts. FITC-labeled tracer was added to culture media in the upper compartment. Following 30 min of incubation, the transmission of tracer from the upper compartment to the lower compartment was analyzed by spectrometry. (B) JAM1 expression in IHGE cells stably expressing shLuc, shJAM1 #110, or shJAM1 #508 was analyzed by immunoblotting with the indicated antibodies. (C–I) Permeability to 40 kDa FITC-dextran (C), 3–5 kDa FITC-dextran (D), 0.5 kDa Lucifer Yellow (E), FITC–P. gingivalis LPS (F), FITC–P. gingivalis PGN (G), FITC–E. coli LPS (H), or FITC–S. aureus PGN (I) in IHGE cells expressing shLuc and shJAM1. Results are expressed as fold change relative to cells expressing shLuc and are the means ± SD of eight technical replicates. *, p<0.05, two-tailed Dunnett’s test (C-G) or two-tailed t test (H, I).