Immunolocalization of SgIGSF in CMCs.

Immunocytochemical analysis of B6^+/+ (A) and B6^tg/tg (B) CMCs. Cytospin preparations of CMCs were fixed with methanol, reacted with the anti-SgIGSF antibody, and stained with FITC-labeled secondary antibody. Bar, 10 μm. (C) Immunocytochemical analysis of the coculture of B6^+/+ CMCs and NIH/3T3 cells. CMCs were cocultured on the monolayer of NIH/3T3 cells for 3 hours. The coculture was fixed with methanol, reacted with the anti-SgIGSF antibody, and stained with FITC-labeled antibody. A representative set of X-Y and Y-Z sections is shown. A red line indicates the plane of the Y-Z section. (D-F) Colocalization of SgIGSF with polymerized actin filaments in the peripheral margin of B6^+/+ CMCs that have adhered to NIH/3T3 cells. After CMCs were cocultured on the monolayer of NIH/3T3 cells for 3 hours, the coculture was fixed with paraformaldehyde, reacted with the anti-SgIGSF antibody, and stained with FITC-labeled secondary antibody (D). Subsequently, the culture was stained with TRITC-labeled phalloidin (E) and the FITC and TRITC images were merged (F). *B6^+/+ CMCs; #NIH/3T3 cells. Bars, 10 μm.