Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions
Figure 1
VSV N, P and L proteins localize to inclusions in infected cells.
(A) CV-1 cells were infected with VSV-eGFP-P at an MOI of 3 and fluorescent microscopy images acquired at 5 hours post infection (hpi). (B) CV-1 cells were transfected with 3.8 µg of a plasmid expressing eGFP-P and examined by fluorescence microscopy at 24 hours post transfection. An image of a single representative cell illustrating the typical distribution of P is shown. (C and D) BSR-T7 cells were infected with VSV-eGFP-P or VSV-RFP-P respectively as in panel A, and N (panel C, red) and L (panel D, green) were detected by immune fluorescence microscopy. (E and F) Vero cells were infected with rVSV at an MOI of 5, and the distribution of the N (red) and L (green) proteins (panel E) or M (green) and L (red) proteins (panel F) was detected by IF microscopy at the indicated hpi. (G) Vero cells were infected with rVSV as in panel A and at 6 hpi were prepared for thin-section electron microscopy. To preserve intracellular membrane structures, samples were fixed with glutaraldehyde as in methods. (H–J) Vero cells were infected with rVSV (H, J) or rVSV-eGFP-P (I) and prepared at 6 hpi for immuno gold labeling as described in methods. Size bars are 5µm (panels A–D), 10µm (panels E, F) and 0.5µm (panels G–J). Note samples B–F were also stained with DAPI to visualize the nuclei. ER = endoplasmic reticulum, Nuc = nucleus, Mit = mitochondria, VIB = viral inclusion body.