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. 2012 Sep 20;8(9):e1002935. doi: 10.1371/journal.ppat.1002935

Figure 8. Glycoprotein analysis of MuHV-4 propagated in RAW-264 cells.

Figure 8

a. EF1α-eGFP virions propagated in BHK-21 (BHK) or RAW-264 (RAW) cells were assayed for virion components by immunoblotting. gB (C) and gB (N) are the C-terminal and N-terminal halves of gB after its cleavage by furin. The upper band corresponds to uncleaved gB. b. Wild-type MuHV-4 virions propagated in BHK-21 cells were bound to NMuMG cell monolayers (3 h, 4°C, 3 p.f.u./cell), then washed ×2 in PBS and either fixed immediately, or incubated further (2 h, 37°C) before fixation to allow virion endocytosis. The cells were then analysed by immunofluorescence for virion antigen display. c. EF1α-eGFP virions propagated in BHK-21 or RAW-264 cells were bound to NMuMG cell monolayers (3 h, 4°C, 25 eGFP units/cell - equivalent to 3 p.f.u./cell), then washed ×2 in PBS, fixed and analysed by immunofluorescence for virion antigen display. Each dot corresponds to a positive virion. The numbers show mean ± SD positive virions per cell for 25 cells per sample. Comparison by Student's t test showed that significantly more RAW-264-derived virions than BHK-21 cell-derived virions were recognized by mAbs MG-1A12 and MG-9B10, and significantly fewer by mAbs T2C12 and 47-5G10 (p<0.01). Equivalent data were obtained in 2 further experiments.