Subcellular localization of JAM members to epithelial cell-cell contacts.

(A) MDCK cells stably transfected with cDNA encoding murine JAM-1, JAM-2, or FLAG-tagged JAM-3 were stained for ZO-1 in red and with antibodies to JAM-1 (H202-106), JAM-2 (XVIIIF26), or FLAG peptide (M2) in green. Confocal images taken at the apical level of tight junctions and micrographs showing the Z-axis reconstitutions are shown. The latter were obtained from a stack of 40 pictures every 0.4 μm taken at the level of the arrowheads. Magnification × 630. (B) MDCK cells expressing EGFP fusion proteins for JAM-1 (i and iv), JAM-2 (ii and v), or JAM-3 (iii and vi). Staining with anti–ZO-1 followed by antirat Texas Red (iv-vi) are shown below the pictures obtained for green fluorescence (i-iii). Micrographs were acquired using specific filter sets and Openlab software. Magnification × 1000.