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Microparticle-Mediated Transfer of the Viral Receptors CAR and CD46, and the CFTR Channel in a CHO Cell Model Confers New Functions to Target Cells

Figure 9

CFTR channel activity

in MP-transduced CHO recipient cells. (A), Evolution of the DiSBAC2(3) fluorescence signal. The time-course analysis of DiSBAC2(3) fluorescence changes was monitored in regions of the cell monolayer corresponding to 20–30 cells, taken at 72 h post-transfer, a time point corresponding to the maximal immunoreactivity of CFTR glycoprotein at the cell surface. Changes in the fluorescent signal were expressed as Ft/F0 ratio values, in which Ft and F0 were the fluorescence values at the times t and t0, respectively, and t0 the time when the cAMP-containing, CFTR-activating cocktail was added. The cocktail of CFTR activators was maintained throughout the experiment. The CFTR inhibitor GlyH-101 was added for 5 min (phase II, marked by two vertical arrows). Phase III shows reversibility of the CFTR block and recovery of the fluorescent signal. (B), Fluorescence microscopy. Photographs of cell monolayers were taken at different time-points corresponding to the four successive phases, as indicated on the curve by the letters (a), (b), (c) and (d).

Figure 9