Expression of <i>M</i>. <i>ulcerans</i> proteins by VRP-infected cells.

<p>(A) Genome maps of recombinant VSV: The genome of VSV encodes for the nucleoprotein N, phosphoprotein P, matrix protein M, glycoprotein G, and the large RNA polymerase L. In VSV*ΔG, the glycoprotein G coding region was replaced by the coding sequence for eGFP. In VSV*ΔG(MUL22332) and VSV*ΔG(MUL3720), the VSV G gene was replaced by the <i>M</i>. <i>ulcerans</i> genes MUL2232 and MUL3720, respectively. The reporter protein eGFP is encoded by an additional transcription cassette located downstream of the <i>M</i>. <i>ulcerans</i> genes. (B) BHK-21 cells were infected with the indicated replicons and indirect immunofluorescence analysis performed with mAbs specific for MUL2232 (B1) or MUL3720 (B2) (red fluorescence). The infection of cells with VRPs is indicated by eGFP expression (green fluorescence). Scale bar equals 30 μm. (C) Equal numbers of L929 fibroblasts were infected with the indicated VRPs at a MOI of 10. At 6 hours post infection, the cells were harvested and protein lysates of the soluble (s) and insoluble (i) fraction analysed by Western blotting for expression of <i>M</i>. <i>ulcerans</i> antigens using mAbs specific for MUL2232 (C1) and MUL3720 (C2), respectively.