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. 2015 Jun 17;11(6):e1004957. doi: 10.1371/journal.ppat.1004957

Fig 9. pUL31-hbpmp1mp2 forms mature nucleocapsids delayed in nuclear egress.

Fig 9

Vero cells were infected with HSV1(17+)Lox or Lox-UL31-hbpmp1mp2 at an MOI of 1, fixed at 13 or 17 hpi and analyzed by electron microscopy. (A-D) Electron micrographs of Lox-UL31-hbpmp1mp2 at 13 hpi revealed all intracellular stages of virus maturation: capsids in the nuclear matrix (A, arrowheads), capsids traversing the nuclear envelope (B, arrowhead), early stages of secondary envelopment (B, white arrow), and fully enveloped virions in vesicles (A-C, black arrows). At the extracellular surface of the cells, virions (D, black arrow) and L-particles (D, arrowheads) were present. (E-G) Quantitation of the number of capsids in the nucleus and the cytoplasm after infection with HSV1(17+)Lox or Lox-UL31-hbpmp1mp2 for 17 hpi; (E) number of intracellular capsids, (F) number of cytoplasmic capsids, (G) ratio of nuclear to total cellular capsids. 12 cells of each condition were analyzed. (E, F) Each data point corresponds to one cell. (G) The ratio of the number of nuclear capsids per mm² and the number of cellular capsids per mm² was calculated and expressed as arithmetic means. (E-G) SDs are indicated. Significance was calculated with an unpaired t test using the software Graphpad Prism (ns = not significant, p = 0.6196; **, p = 0.0043; ***. p<0.0001). Scale bars equal 200 nm.