Figure 7.

The effect of unlabelled Q44c, Q86c and EGF on the binding of fluorescent (A) Q44c-HL488 (14.6 nM) and (B) Q86c-HL488 (12.5 nM) to full-length N-terminal nanoluciferase-EGFR. NanoBRET experiments were performed using NLuc-EGFR stably expressing HEK293 cells. Cells were treated with either nanobody and EGF simultaneously and incubated for 30 minutes at 37^oC. The NLuc substrate furimazine (12.5 nM) was added and plates incubated for 5 minutes then luminescence and fluorescence emissions were measured using a BMG Pherastar. Experiments were performed in HBSS containing 0.2 % BSA. Data are combined mean ± SEM from five independent experiments, where each experiment was performed in triplicate.