Nipah virus induces two inclusion body populations: Identification of novel inclusions at the plasma membrane
Fig 2
Colocalization of NiV M protein and inclusion bodies in NiV-infected and transfected cells.
(A) Vero76 cells were infected with wildtype NiV and NiVΔM at a MOI of 0.01 and 0.025, respectively. At 24 h p.i. cells were fixed with 4% PFA for 48 h and permeabilized with Triton X-100. Cells were incubated with a NiV N-specific guinea pig antiserum to visualize IBs (green). NiV M was detected with an M-specific rabbit antiserum (red). Nuclei were counterstained with DAPI (blue). Confocal xy sections of the central regions of syncytia are shown. White dotted lines indicate the lateral borders of syncytia. (B) Cells coexpressing the NiV proteins N, P, G and F in the presence (N/P/G/F + M) or absence of NiV M (N/P/G/F) were fixed and permeabilized with Triton X-100 at 24 h after transfection. Immunostaining was performed as described above. Scale bars, 10 μm. In the right panels, quantifications of IB distribution in syncytia is shown. Using the automated ImageJ analyze particle tool, the total numbers of IBs and the number of IBs located at a maximum distance of 10 μm from the lateral edge of the syncytium were counted in individual sections of 6–10 syncytia from three individual experiments. The average percentage of membrane-proximal IBs in syncytia in the absence and presence of M was calculated. Error bars indicate the standard error of the mean. Statistical significance is indicated by asterisks (unpaired t-test; ***, p < 0.001).