NCAM180 induces Ric8A-GFP recruitement at the membrane where NCAM and Ric8A colocalize.

<p>A- Ric8A-GFP co-transfected with NCAM120, NCAM140 or NCAM180 in HEK293T cells co-localizes specifically with the NCAM180 isoform. Scale bar: 10 µm. B- Ric8A-GFP or PmaxGFP plasmids were transfected in cortical neurons for 24 h. Images were taken focussing at the membrane plane with a 20× objective. NCAM staining is clearly associated to the cell membrane (B, E, K). Ric8A-GFP is strongly localized at the membrane at the level of the cell body and processes (A, D). PmaxGFP control plasmid is mostly found in the cytoplasm (J). Ric8A-GFP, but not PmaxGFP colocalizes with NCAM at the membrane (Ric8A/NCAM merge: C, F versus Pmax/NCAM merge: L). (G–I): higher magnification pictures (dotted line box D–F) using a 63× oil immersion objective indicate a strong co-localization of Ric8A-GFP with NCAM in the growth cone (I). Scale bars: 10 µm (A–F, J–O); 1 µm (G–I).