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. 2013 Jun 10;8(6):e66059. doi: 10.1371/journal.pone.0066059

Figure 1. Expression of functional hemagglutinin (HA) with recombinant VRPs.

Figure 1

(a) Genome maps of recombinant VSV: the parental VSV genome contains five transcription units encoding for the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G), and the large RNA polymerase (L). VSV*ΔG lacks the glycoprotein G gene but contains the eGFP gene instead (the asterisk denotes for eGFP). VSV*ΔG(HA) expresses the influenza virus HA from the fourth gene position while eGFP is expressed from an additional transcription unit downstream of HA. (b) Immunofluorescence analysis of Vero cells 8 hours p.i. with either VSV*ΔG, VSV*ΔG(HAH5-HP) or VSV*ΔG(HAH5-LP). At 5 hours p.i. with VSV*ΔG(HAH5-LP), the cells were treated for 60 minutes with trypsin (+T) or were left untreated (−T). Thereafter, the cells were exposed for 5 minutes to either pH 5.4 or pH 7.4, incubated for 60 minutes at 37°C with normal medium, fixed with formalin, and finally processed for immunofluorescence using a swine antiserum to HAH5 (red fluorescence). Nuclei were stained with DAPI (blue fluorescence). Expression of eGFP is indicated by green fluorescence. Scale bar represents 20 µm.