Construction and Characterization of Insect Cell-Derived Influenza VLP: Cell Binding, Fusion, and EGFP Incorporation
Figure 3
HA-mediated endocytosis of VLP in A549 cells. A549 cells were added to VLPs labeled with DiO dye and incubated on ice for 1 h. The unbound VLPs were washed away, and the cells were incubated at 37°C for further 2 h. (b) and (c) show A549 cells incubated with HA− VLPs or HA+VLPs, respectively. (d) displays VCNA-pretreated A549 admixed HA+ VLP. (a) is a mock control. The upper and right rectangular images of each panel depict horizontal and vertical cross-sections located at green and red lines, respectively. VLPs were labeled with DiO dye (green), and A549 cells were stained with DiD dye (pink). Graph in (e) represents the average of endocytosed VLPs per cell. Graph in (f) depicts the inhibitory effect of fetuin on VLP binding. The values of relative VLP binding was determined by the ratio of R18 signals at each concentration of fetuin to the value obtained for the maximum binding of VLPs without the fetuin treatment.
