Fig. 6. CD82 causes redistribution of EGFR and GD1a. HB2/ZEO (A,C,E) and HB2/CD82 cells (B,D,F-J) were grown on glass coverslips for 48 hours. Cells were fixed with paraformaldehyde and indirect immunofluorescence staining was carried out using mAbs to EGFR (A,B), GD1a (C,D), FITC-conjugated cholera toxin (E,F). Double staining was carried out using a combination of anti-GD1a (GD1a-1) and anti-CD82 (C33) mAbs (G,H) or anti-GD1a (GD1a-1) and anti-EGFR (Ab-16) mAbs (I,J). Staining was visualised using FITC-conjugated goat anti-mouse IgG (A-D) or a combination of Texas Red-conjugated goat anti-IgG1 and Alexa Fluor 488-conjugated goat anti-IgG2a (G-J). H and J are digitally magnified highlighted areas of G and I, respectively (x6). Images were acquired using the Nikon Eclipse E600 microscope (Plan Apo 60xA/1.4 oil) and the Leica DC200 digital camera. The images were subsequently processed using the DC200 image processing programme.

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