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. 2020 Apr 28;10:186. doi: 10.3389/fcimb.2020.00186

Figure 1.

Figure 1

US28 protein expression is ablated in stopUS28-infected fibroblasts. (A) BFXwt-GFP (wt) was used to generate inUS28-3xF, which contains an in-frame triple FLAG epitope tag (3xF) at the C-terminal end of the ORF (checked arrow). inUS28-3xF was then used as the template to generate two independent stop mutants, stopUS28-1 and stopUS28-2, that each contains a stop codon following the first methionine (red stop sign). (B,C) Fibroblasts were infected as indicated (moi = 0.5). (B) Cell lysates were harvested 96 hpi for immunoblot. α-FLAG was used to detect pUS28 expression, α-pp65 is a marker of infection, and actin is shown as a loading control. (C) Infected and mock cultures were processed for IFA 72 hpi. α-FLAG was used to detect pUS28 expression via the 3xF epitope (red). eGFP (green) is a marker of infection, and nuclei were visualized using DAPI (blue). Images were acquired using a 60x objective. (B,C) Representative images are shown; n = 3.