Supplementary Material

JCS072058 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Double immunofluorescence staining of frozen sections of mouse epididymis, small intestine, and liver with anti-angulin/LSR pAb (green) and anti-occludin mAb (red). Three independent experiments showed similar results. Junctions of linear occludin staining indicate tricellular contacts. In epididymis, all of the 164 tricellular contacts are positive with LSR concentration. In the small intestine, 231 of 233 tricellular contact are positive with LSR concentration. To score the exact number of tricellular contacts is difficult in the liver due to the complex organization of the cell-cell junctions at bile canaliculi, but many LSR concentrations are observed at the junctions of occludin staining. Bar, 20 µm.

• Supplemental Figure S2 -

  Fig. S2. Double immunostaining of cocultured GFP-tagged angulin/LSR-expressing L cells and HA-tagged angulin/LSR-expressing L cells with anti-GFP mAb (green) and anti-HA mAb (red). GFP-tagged angulin/LSR and HA-tagged angulin/LSR are often colocalized (arrowheads). Bar, 10 µm.

• Supplemental Figure S3 -

  Fig. S3. Immunofluorescence staining of subconfluent EpH4, KD1 and KD1-resc1 cells with anti-occludin mAb. Bar, 20 µm.

• Supplemental Figure S4 -

  Fig. S4. (A) Double immunofluorescence staining of L cells (−) stably expressing Tric-full and GFP-angulin/LSR-L cells stably expressing Tric-full, Tric-ΔN or Tric-ΔC with anti-GFP mAb (green) and anti-HA mAb (red). In the merged images, DAPI-stained nuclei (blue) are also shown. (B) Double immunofluorescence staining of GFP-angulin/LSR-L cells stably expressing CD9 or CD9-TricC with anti-GFP mAb (green) and anti-HA mAb (red). In the merged images, DAPI-stained nuclei (blue) are also shown. Bars, 10 µm.