JCS00857 Supplemental Data

Files in this Data Supplement:

• Supplement Figure1 -

  Fig. S1. Cadherin adhesions are formed in HeLa and MDCK cells. N-cadherin-GFP transfected HeLa (a) or MDCK (b) cells were plated onto Ncad-Fc substrate in the same experimental conditions as C2 cells (described in Materials and Methods). HeLa cells were directly observed for GFP under video-microscopy after 30 minutes of spreading.  MDCK cell were fixed after 8 hours of spreading in the absence of serum and stained for b-catenin. More than 90% of attached cells on the Ncad-Fc substrate expressed N-cadherin-GFP, in agreement with the homophilic properties of the ligand. Living HeLa cells displayed radial structures of N-cadherin in the lamellipodium. MDCK cells present a similar pattern of b-catenin, although radial structures were notably shorter and accumulated in a reduced lamellipodium. Bar: 10 mm.

• Supplement Figure2 -

  Fig. S2. Endogenous Rac1, but not PI 3-kinase organised in radial structures in the lamellipodium. C2 cells were allowed to attach on Ncad-Fc substrate for 2 hours, fixed by cold methanol (A) or formaldehyde (B) then stained for endogenous Rac1 and b-catenin (A) or PI 3-kinase (B, antibody directed against the p110 subunit, Calbiochem, 1/200 dilution). (A) The endogenous Rac1 immunostaining partially accumulated in b-catenin-containing radial structures. In addition, it was enriched at the tip of the lamellipodium and in the ruffles, formed by this cell. (B) The PI 3-kinase immunostaining uniformly distributed in the lamellipodium, with some accumulations at the leading edge. Bar: 10 mm.