Figure 3. Intracellular [Ca²⁺] increases during PRV infection in neurons and is dependent on gB-dependent membrane fusion.

(A and B) Mock and infected neurons were stained with the Ca²⁺-sensitive dye OGB-1. Neurons were imaged every 10 minutes between 3–22 hpi. (A) Change in fluorescence intensity for individual neuron cell bodies over the course of infection (following subtraction of background fluorescence intensity). Bolded lines represent the average change in fluorescence intensity for each condition, while light lines show trends for 8 individual representative neurons for each condition (total data is n=22 for mock, n=38 for PRV Becker, n=18 for PRV gB-null, acquired from neuron cell bodies in two fields of the same dish; additional biological replicates are presented in Figure S2A). (B) Representative images of intracellular Ca²⁺ in mock and PRV infected neurons between 3–18 hpi. Phase contrast and OGB-1 staining (green) are shown (scale bar = 50 µm). (C) Quantification of the percentage of motile mitochondria in axons from PRV Becker infected SCG neurons at 16–20 hpi. Neurons were treated with 2mM EGTA (added at 0 hpi) or 150 µM CdCl₂ (added at 15 hpi). Error bars represent mean ± SEM. See also Figure S2 and Movie S2.