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Fig. 2. Recombinant IgLON-Fc proteins show differential interactions with CGCs. CGCs adhere preferentially to CEPU-1-Fc and fail to adhere to LAMP-Fc. Dissociated CGCs were incubated on coverslips sequentially coated with protein A and recombinant IgLON-Fc for (A) 4 hours (n=2) or (B) 18 hours (n=3), and adhering cells were counted. After 4 hours, the number of cells adhering to CEPU-1-Fc was approximately 60% of that adhering to poly-L-lysine and was significantly above background (P<0.0001), whereas adhesion to LAMP and OBCAM was similar to background. After 18 hours, adhesion to CEPU-1-Fc had increased to 80% of the poly-L-lysine control and adhesion to OBCAM-Fc was now 50% of the poly-L-lysine control and significantly above background (P<0.0001). Adhesion to LAMP-Fc remained similar to background. (C) Live staining of CGC with IgLON-Fc revealed a punctate stain with both CEPU-1-Fc and OBCAM-Fc, but LAMP-Fc staining was similar to the secondary antibody alone. In keeping with the adhesion results shown above, the CEPU-1-Fc staining was more intense than that observed with OBCAM-Fc. Bar, 25 µm.
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