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. Author manuscript; available in PMC: 2017 Jan 1.
Published in final edited form as: Virology. 2015 Oct 19;487:19–26. doi: 10.1016/j.virol.2015.10.001

Figure 6.

Figure 6

Crm1 inhibition. (A) Leptomycin B (LMB, 4 nM) was pre-incubated with rCrm1 before the addition of bait proteins (singly phosphorylated GSL-LE or GST) and (putative) auxiliary prey proteins (rRan with RCC1). The complexes were extracted with glutathione-sepharose beads, fractionated on gels and detected by Western analyses. (B) Plated HeLa cells were pre-incubated with LMB or its carrier (MeOH) before infection with vEC9 at the indicated MOI. The cells were harvested at 4 hr PI. Nup62 phosphorylation levels (% intensity) were determined by Western analyses and densitometry relative to samples with no inhibitor or virus, and as normalized to each lane’s tubulin levels. (C) Plates of confluent HeLa cells were infected with vEC9 (MOI of 10) after pretreatment (or not) with LMB. After 6 hrs, recovered progeny virus was titered by plaque assay. Observed PFU/ml averaged from 3 independent experimental iterations are indicated (SD: 0.44–0.49).