Fig. 2

Internalization of BILF1 receptors depends on dynamin-1 and clathrin. SNAP-tagged EBV-BILF1 (green), SNAP-tagged PLHV1-BILF1 (blue), SNAP-tagged PLHV2-BILF1 (orange), and empty vector (gray) were expressed in HEK-293A cells and were either a co-transfected with different concentrations of Dyn K44A DNM (black circle 3 ng; black up-pointing triangle 7 ng; black diamond 11 ng) or c incubated with 5 µM Pitstop2 inhibitor (black circle or white circle for empty vector) or internalization buffer as a control (black square). Labeling with donor SnapLumi4-Tb was performed at 4 °C to prevent internalization prior to measurement. Internalization was measured every 4 min for 88 min at 37 °C. The graph curves represent the ratio between donor and acceptor. b, d The area under the curve (AUC) was calculated for each graph and was normalized to the AUC of each BILF1 receptor without addition of the Dyn K44A or Pitstop2 (100%) and empty vector (0%). Statistical differences were determined using Dunnett’s multiple comparisons two-way ANOVA test. *P < 0.05, **P < 0.01, *** P < 0.001, ****P < 0.0001