Fig. 2

Pre-treatment effect of RES on PHEV replication in N2a cells. A Schematic diagrams illustrating the experimental design for time-of-addition experiments. B, C Expression of PHEV protein and mRNA in N2a cells at 24 h as determined by Western blot and qPCR assay, respectively. PHEV protein and mRNA levels were quantified and normalized to GAPDH levels, respectively. D TCID₅₀ values were the means of three repeated titrations at the time points indicated. E An immunofluorescence assay was performed to examine PHEV expression in N2a cells at 24 h after RES treatment. Labeled and stained cells were viewed using confocal microscopy. PHEV-positive cells were labeled with an anti-N protein monoclonal antibody (green) and nuclei were stained with Hoechst (blue). Scale bar = 100 μm. Data are presented as means ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 based on comparisons to untreated PHEV-infected cells