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. 2016 Jun 9;6:27546. doi: 10.1038/srep27546

Figure 4. Fiber codon optimization in the adenovirus interferes with the expression of the armed protein when expressed under the control of the Major Late Promoter (MLP).

Figure 4

(A) Schematic representation of the Adwt-EGFP and AdFO-EGFP genomes with the EGFP engineered as an L6 gene under the control of the MLP. (B) EGFP values were analyzed by fluorescence quantification 30 h post-infection in A549 cells infected with Adwt-EGFP or AdFO-EGFP. Left panel shows representative images of EGFP expression. Right panel shows EGFP fluorescence intensity quantification of 20 cells per experiment of three independent experiments shown as a mean ± SEM. (C) Representative western blot of hexon and fiber proteins 30 h post-infection. (D) Intracellular and extracellular viral DNA content analyzed at 30 h post-infection. (E) Comparison of extracellular viral DNA release between AdFO and AdFO-EGFP, versus Adwt and Adwt-EGFP respectively analyzed by qPCR. Cells were infected using 10 TU/cell of both viruses. Data is shown as mean ± SEM of five independent experiments. All AdFO-EGFP fluorescence, intracellular DNA and viral release values are expressed as relative to their corresponding value for Adwt-EGFP in each replicate. ***p < 0.01, ***p < 0.001.