Fig. 3. A novel experimental system to quantitatively determine FGF-2 export from
living CHO cells. CHOFGF-2-GFP cells were grown for 18 hours at
37°C under the conditions indicated followed by dissociation from the
culture plates by using a protease-free protocol. The cell suspension was then
processed for the FACS analysis under the conditions indicated. Panels A-E
represent dot blots where total GFP-derived fluorescence was blotted against
cell surface-derived PE fluorescence. (A) Cells grown in the absence of
doxicyclin. (B) Cells grown in the presence of doxicyclin and processed with
anti-GFP antibodies. (C) Cells grown in the presence of doxicyclin and
processed with anti-GFP antibodies. (D) Cells grown in the presence of
doxicyclin followed by a wash procedure using a heparin-containing buffer and
antibody processing. (E) Cells grown in the presence of doxicyclin followed by
trypsin digestion and antibody processing. Panels F and G represent the
corresponding histograms of GFP-derived fluorescence and PE-derived cells
surface fluorescence, respectively. The colours correspond to the conditions
shown in panels A-E.