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. 2012 Apr;86(8):4182–4193. doi: 10.1128/JVI.07105-11

Fig 4.

Fig 4

Mutations in the EIAV-PTAP (ePTAP) NC domain interfere with virus budding. Coexpression of EIAV L domain mutant eYP−, which harbors a functional NC, rescues the release of ePTAP NC mutants ePTAP-RKI (A) and ePTAP-RKII (B). 293T cells were transfected with ePTAP (lane 1), eYP− (lane 2), or ePTAP-RKI or -RKII (lane 3) alone or cotransfected with the indicated NC mutant and eYP− plasmids (lanes 4 and 5). DNA ratios of the indicated NC mutant to the YP mutant used were 4:1 and 1.5:1, while the total amount of DNA remained constant. Viral fractions and cell lysates were analyzed by Western blotting using the indicated antibodies. Relative virus release efficiencies are shown at the bottom (± the standard deviations; n = 3). (C) ePTAP NC mutants displayed typical budding defects in 293T cells. Shown are transmission EM images of thin-sectioned 293T cells transfected with ePTAP-WT (a), ePTAP-RKI (b and d), or ePTAP-RKII (c and e). Arrows indicate either WT mature virions (a) or tethered arrested particles (b to e). Scale bars, 200 nm.