Fig 6.

Effect of SDC-1 plasmid transfection, antibody-mediated SDC-1 inhibition, and SDC-1 recombinant protein competition on baculovirus transduction and binding. (A) HepG2 cells were pretreated with SDC-1-specific (0, 10, and 20 mg/ml) and control antibodies (20 μg/ml; IgG). Baculovirus (MOI, 800) was added, and the percentage of EGFP-positive cells was analyzed 48 h later by FACS. (B) HepG2 cells were pretreated with 20 μg/ml of SDC-1 antibody. Baculovirus (MOI, 800) was added and allowed to enter the cells for 30 min at 37°C. Baculovirus was stained with anti-vp39 and anti-mouse Alexa 488 antibodies, and SDC-1 was stained with anti-rabbit Alexa 594 antibody. Baculovirus is seen in green and SDC-1 in red. Imaging was performed by confocal microscopy (20× magnification). (C) HepG2 cells were pretreated with SDC-1 recombinant protein (0, 1, 2.5, and 5 μg/ml) and the baculovirus (MOI, 200) was added. The percentage of EGFP-positive cells was analyzed 48 h later with FACS. (D) HepG2 cells were transfected with SDC-1-encoding plasmid, and baculovirus (MOI, 400) was added 24 h later. The percentage of EGFP-positive cells was analyzed 48 h later by FACS. In all transduction experiments, EGFP/WPRE-bearing baculovirus was used. Mean fluorescence values and standard deviations are shown.