Figure 3.

EspF and Map prevent the junctional recruitment of proteins during de novo junction assembly. EspF and Map, but not Tir, prevent the incorporation of proteins in forming junctions. Cells were cultured on cover-slips in low calcium (<5 µM) medium for 18–20 hours and then switched to normal calcium (~1.8 mM) medium. After 14 hours, cells on cover-slips were fixed and stained with primary antibodies against the indicated tight junction proteins and Cy3-conjugated secondary antibodies. Nucleus was labeled with DAPI (4′,6-Diamidino-2-phenylindole dihydrochloride). Cover slips were mounted on glass slides and images were acquired on a Zeiss ApoTome (Axiovert 40 CFL) using 63X oil objective. (n = 3); scale: 10 µm.