Fig. 6. Homophilic cell-cell adhesion activity of Necl-1. (A) Cell aggregation activity of Necl-1 when a single-cell suspension was rotated for 10 minutes. Aa, wild-type L cells; Ab, Necl-1-L cells. Bars, 100 µm. (B) Localization of Necl-1 at cell-cell contact sites of Necl-1-L cells. Necl-1-L cells were stained with anti-FLAG mAb. Arrow indicates cell-cell contact sites. Bar, 10 µm. (C) Homo-cis-dimer formation of Necl-1. A single-cell suspension of Necl-1-L cells was incubated in the presence or absence of BS3. The lysates (20 µg of protein each) were subjected to SDS-PAGE (10%), followed by western blotting with anti-FLAG mAb. Arrow, monomer; arrowhead, dimer; double-arrowhead, multimer. The results shown are representative of three independent experiments.