FIG 5.

Infectivity of HAstV-8 in cells transfected with Dyn2 and Eps15 dominant-negative mutants. (A) Caco-2 cells were transfected with the indicated constructs as described under Materials and Methods and incubated for 48 h, at which time they were infected with trypsin-treated HAstV-8. Fourteen hours p.i., the cells were fixed and immunostained with anti-HA (green; to detect the expression of Dyn2) and with anti-HAstV-8 (red) antibodies. (B) Cells expressing Eps15 mutants were detected by the fluorescence of the fused GFP. Cells transfected with the wild-type proteins and also infected are marked with arrows, whereas cells transfected with the DN mutants that are not infected are marked with dotted lines. (C) At least 300 cells transfected with the indicated plasmids were screened for HAstV-8 infection, and the numbers are expressed as percentages of the infected cells observed in nontransfected cultures. The arithmetic means and standard deviations of three independent experiments performed in duplicate are shown. The images were obtained by confocal microscopy with slices of 0.2 μm.