Fig. 2

Mutant-VSV effectively infects neurons in vivo. a Rescue and preparation process of VSV. BHK-21 cells were pretreated with vaccinia carrying T7 RNA polymerase for one hour, then washed with PBS three times, and co-transfected with VSV genome plasmid and packaging plasmids encoding the N, P and L proteins. After 6 h, they were replaced with fresh culture medium and placed in a 31 °C incubator. 5 days later, the supernatant was collected and filtered with 0.1 μm filter membrane to remove the vaccinia, then added to BHK-21 cells to amplify the VSV. b Fluorescent imaging of BHK-21 cells infected with WT-VSV and Mutant-VSV at an MOI of 0.001. When infected with WT-VSV, obvious fluorescence signals can be detected within 12 h, while Mutant-VSV needs 24 h. c The single-step growth curves of the WT-VSV and Mutant-VSV. The viruses were collected and titered on BHK-21 cells at indicated time points (12, 24, 36, 48, 72 and 96 hpi). d Fluorescent signals from mutant VSV were co-localized with neurons in vivo. Cell nuclei were stained using DAPI (blue), neurons were stained with Cy3-conjugated NeuN antibody. Scale bars =100 μm for B and D