FIGURE 5.

Hyper-triggering HN mutant with mutation in site II at HN dimer interface (H552Q) shows enhanced interaction with F. A, 293T cells co-transfected with constructs encoding HN wt N-Venus and F C-CFP or HN H552Q N-Venus and F C-CFP were treated overnight with 10 mm zanamivir, and 1 h before analysis fresh 10 mm zanamivir and cycloheximide were added. Evenly distributed fluorescence is visualized across the cell surfaces in both representative fields. The immunoblot shows the expression level of HN and F for HN wt N-Venus, F C-CFP and HN H552Q N-Venus, F C-CFP. Anti-tubulin (55 kDa) loading controls are below. B, shown is the mean fluorometric ratio resulting from HN-F interaction observed in panel A, measured in an average of at least three fields per experiment, from seven separate experiments. ****, p < 0.0001 (one-tailed t test).