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. 2018 Feb 24;10(2):95. doi: 10.3390/v10020095

Figure 3.

Figure 3

Two divergent strains of ZIKV can both cleave the ZIKV-NLS-GFP reporter. (a) Alignment of viral amino acids adjacent to the NS2B-NS3 cleavage site (amino acids 2511–2530 of ZIKV and 2482–2501 of DENV-2), including the Puerto Rico (ZIKV-PR) and Dakar (ZIKV-DAK) ZIKV strains, as well as dengue virus serotype 2 (DENV-2). (b–d) Immunoblot analysis of extracts from A549 cells stably expressing ZIKV-NLS-GFP that were mock-infected or infected with an increasing MOI (1, 5, or 10) of ZIKV-PR (b), ZIKV-DAK (c), or DENV-2 (d). Arrows mark full-length (FL) or cleaved (C) ZIKV-NLS-GFP reporter. (e) Confocal micrographs of A549 cells stably expressing the ZIKV-NLS-GFP reporter (green) that were infected with DENV-2 (MOI 10) for 48 h and then immunostained for DENV-2 (anti-Envelope (Env); red). Nuclei were stained with DAPI (blue). Scale bar, 10 µm. (f) The ratio of nuclear to total fluorescence intensity, as determined using Image J, in uninfected and DENV-2-infected cells immunostained for DENV Env. Each dot represents an individual cell (n = 18). Dot plots represent the mean ± SD. (ns) denotes p > 0.05, as determined by an unpaired t-test.