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. 2008 Apr;172(4):905–915. doi: 10.2353/ajpath.2008.070698

Figure 2.

Figure 2

Cldn-153–80 binds to T84 cells and induces TJ disassembly. A: Confluent T84 monolayers were incubated in either control medium (a–e) or in the presence of 200 μmol/L Cldn-153–80 (f–j) for 24 hours and the effect on localization of claudin-1 (a,f), occludin (b,g), ZO-1 (c,h), JAM-A (d,i), and E-cadherin (e,j) was assessed by immunofluorescence confocal microscopy. Cldn-153–80 treatment altered the localization of the TJ proteins (f–i), but not E-cadherin. Bar = 10 μm. B: Cells were incubated in either control medium (a,e,i) or in the presence of 200 μmol/L *Cldn-131–53 (b,f,j), *Cldn-153–80 (c,g,k), or *Cldn-1146–160 (d,h,l) for 24 hours. The cells were then washed, UV-crosslinked, fixed, and treated with FITC-streptavidin to label bound peptide (a–d) and Alexa588-phalloidin to label F-actin (e–h). Merged images are in (i–l). Only *Cldn-153–80 showed high levels of binding to the cells, while the other peptides only show occasional cell association (arrows). Scale bar = 10 μm. C: Cells were either incubated in control medium (a) or in the presence of *Cldn-153–80 (b) for 24 hours and then processed for avidin gold electron microscopy. Black arrowheads indicate abundant association of *Cldn-153–80 with the cells, arrows show internalized *Cldn-153–80 and white arrowheads indicate TJs. Altered TJ morphology was observed in cells treated with *Cldn-153–80 as compared to untreated cells. Scale bar = 667 nm.