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. 2012 Oct 16;4(10):2137–2161. doi: 10.3390/v4102137

Figure 3.

Figure 3

D471G substitution affects the ability of LCMV-NP to promote replication and gene expression of an LCMV MG. BHK-21 cells were co-transfected in triplicate with 0.5 μg of pPolI GFP-Pur/Gluc, together with 0.6 μg of pC-L, 0.3 μg of pC wt or mutant NPs HA-tagged, and 0.1μg of the pSV40-Cluc expression vector to normalize transfection efficiencies At 48 hpt, MG driven GFP expression (A) and luciferase activity in TCS (B) were determined. Cell lysates were used to determine expression levels of wt and mutant NPs by WB using an anti-HA antibody (C). GAPDH was used as a loading control. Empty pC was used as a negative control. Percentages of relative luciferase units (% RLUs) were normalized with respect to the activity of the wt NP, after normalization of transfection efficiencies based on Cluc luminescence values. Numbers at the bottom of each WB lane represent the quantification of band intensities normalized to wt NP lane as described in material and methods.