Skip to main content
. Author manuscript; available in PMC: 2015 May 1.
Published in final edited form as: Virology. 2014 Apr 3;0:96–107. doi: 10.1016/j.virol.2014.03.017

Figure 3. Efficiency of identified NS1 amino acid substitution proteins to trans-complement WNV ΔNS1 GFP VRPs.

Figure 3

(A) HeLa-NFκB-luc reporter cells were transduced with lentivirus expressing either wt NS1, MCS, or single or double NS1 amino acid changes and selected for puromycin resistance. 5μg of whole cell lysate from each cell line was probed for NS1 expression by immunoblot and normalized to β-actin expression. (B) HeLa-NFκB-luc reporter cells stably transduced with lentivirus expressing either wt NS1, MCS, or single or double NS1 amino acid substitutions were infected at an MOI of 0.15 for 36h with WNV ΔNS1 GFP VRPs or mock infected. Expression of GFP was quantified by flow cytometry by determining the mean fluorescent intensity (MFI). The background MFI of identically infected MCS cells was subtracted from the MFI determined for other infected cells. Data are the average of two independent experiments and error bars represent standard deviation from the mean. (C) GFP expression due to trans-complementation by wild type and NS1 proteins with coding changes was monitored by fluorescence microscopy.