Identification of BoHV-5 infection. Detection of BoHV-5 antigens, viral DNA and glyco-C m RNA expression in infected bWJ-MSCs differentiated into neuron-like cells. A) Intense fluorescence positive signal for BoHV-5 glyco-C protein at 72 h post-infection (arrow head); B) Positive grains observed in the neuron-like cell’s nucleus, outside the nuclear membrane and along the axon branches (arrows head) using an in situ hybridization assay; C) Viral growth curve showing a peak of BoHV-5 antigens and cytopathic effect detection at 72 h p.i.; D) Percentage of neuron-like cells positively labeled for viral DNA at all p.i. time points; E) Real-time PCR was used to quantify the mRNA levels and fold-changes were calculated by 2 -ΔΔCt method as compared with expression of endogenous bovine GAPDH mRNA level to normalization. The infected neuron-like cells revealed glyco-C mRNA synthesis at same levels from 24 h to 120 h post-infection (p < 0.05).
Cardoso et al. BMC Veterinary Research 2012 8:242 doi:10.1186/1746-6148-8-242