JCS03233 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Subdomain organization of the strands in a TAJ. Freeze-fracture view of a TAJ at some distance from the tri-cellular contact region shown in Fig. 3a, indicates that the subdomain organization is maintained throughout the DC-OHC junctions. The parallel strands of the claudin-14-enriched region of DC-OHC junctions (upper) are clearly visible and distinct from the meandering strands of the claudin-9/6 enriched region (lower). Bar, 0.1 μm.

• Supplemental Figure 2 -

  Fig. S2. Freeze-fracture view of COS-7 cell plasma membranes. This field shows the random distribution of particles in the plasma membranes of COS-7 cells and demonstrates that these cells do not normally assemble tight junction strands. Bar, 0.15 μm.

• Supplemental Figure 3 -

  Fig. S3. Validation of polyclonal antibodies in transfected COS-7 cells. COS-7 cells transfected with plasmids encoding GFP-claudin-6 (a,b), GFP-claudin-9 (c,d) or GFP-claudin-14 (e-h). GFP fluorescence (green) shows the localization of claudins to contacts between transfected cells (arrows) and antibodies raised against each claudin (red) colocalize extensively. Non-transfected cells (asterisks) do not assemble tight junctions with transfected cells. All cells were counterstained for actin with rhodamine phalloidin (b, d, f and h). Bar, 5 μm.