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. 2018 Oct 24;9:2546. doi: 10.3389/fmicb.2018.02546

FIGURE 5.

FIGURE 5

Effect of single amino acid substitutions in WI/66 NS1 on inhibition of host gene expression. (A) Schematic representation of WT WI/66 (WI WT, gray) and WT HK/97 (HK WT, black) NS1 proteins and the different WI/66 (WI 1 – WI 8) mutant constructs. (B–D) HEK293T cells were transiently co-transfected as described in Figure 1. At 24 hpt, cells were analyzed for GFP (B) and Gluc (C) expression as indicated in Figure 1. ∗P ≤ 0.0001 using One-way ANOVA. NS1 protein expression levels from total cell lysates were analyzed by Western blot (D) using an anti-HA (α-HA) pAb. Actin was included as internal loading control. Molecular mass markers (in kDa) are indicated. Representative images of three independent transfections are represented. Scale bar, 200 μm (B). Data were represented as relative activity considering the activity of HK/97 as 100%. Data represent the mean and SDs of triplicate values (C). Molecular protein markers in kDa are indicated on the right of each of the Western blots (D).