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. 2016 Mar 17;11(3):e0151516. doi: 10.1371/journal.pone.0151516

Fig 1. Ad-mediated FAST protein expression from an E1-deleted vector does not affect virus growth or yield.

Fig 1

A) All viruses are early region 1 (E1) and early region 3 (E3) deleted. AdFAST-HA has a single HA tag linked to the C terminus through a glycine linker. ITR denotes inverted terminal repeats and Ψ is the packaging signal. RFP is the red fluorescent protein and CMV represents the cytomegalovirus enhancer/promoter. B) 293 and A549 cells were infected with the control virus AdEmpty or AdFAST-HA at an MOI of 10. Whole cell lysates were collected 48 hpi. Samples were probed for the HA tag, Ad5 fibre and alpha-tubulin for a loading control. The top band denoted by * in the A549 samples is non-specific. C) 293 cells were infected with AdEmpty or AdFAST at an MOI of 1 and whole cell lysates were collected at 6, 18, 24 and 30 hpi. Samples were probed for Ad5 fibre and alpha-tubulin was used as a loading control. D) Supernatants collected from 293 cells infected with AdEmpty or AdFAST at an MOI of 1 were used to conduct plaque forming assays to determine the number of viral progeny at various times post-infection.