FIGURE 8.
Palmitoylation dependence of gB surface expression and fusion activity in human endothelial ARPE cells. A, plasma membrane steady-state expression (dark gray) and dynamic trafficking (light gray) of gB or gBC777A expressed by mRNA transfection in ARPE cells were measured by CELISA as described under “Experimental Procedures.” Error bars indicate mean ± S.D. Lower panel, anti-gB immunoblot on transfected ARPE total cell extracts. OD, optical density; A. U., absorbance units. B, gB or gBC777A mRNA was co-transfected with or without mRNAs coding for gH pentamer subunits (gH 5) and T7 RNA polymerase (T7RNAP) into ARPE cells and then mixed with cells transfected with p7IRESLuc DNA (see “Experimental Procedures”). gH 5- and T7RNAP-positive, gB-negative transfections were used as control. In one experimental group, the fusion assay was carried out in the presence of 50 μm 2Br-palmitate (2Br-p). Luciferase activity was recorded and plotted after subtracting mock transfection signals. Error bars indicate mean ± S.D.