Fig. 3. EGFP/hSP-C^1-197 chimeric protein is directed to CD63 positive cytoplasmic vesicles. A549 cells grown on glass coverslips at 50-60% confluence were transfected with 10 µg of EGFP-C₁/SP-C^1-197 using CaPO₄ as described in Materials and Methods. At 48 hours after transfection, cells were fixed, permeabilized and stained with primary monoclonal CD-63 antiserum and IgG-specific secondary Texas-Red-labeled antisera. Images were acquired by video fluorescence microscopy as in Fig. 2 and are representative of six separate experiments and >50 cells for each construct. The majority of EGFP/hSP-C^1-197 (A) was associated with CD-63 staining (B). Phase image is shown in C.