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. 2022 Jun 3;18(6):e1010593. doi: 10.1371/journal.ppat.1010593

Fig 1. Generation of a reporter ZIKV that is unable to replicate but is rescued by the exogenous expression of NS1.

Fig 1

(A) The HiBiT luciferase was inserted in the β-ladder of NS1 (NS1 dimer: Protein Data Bank accession number: 4TPL). A schematic representation of the recombinant ZIKV carrying the HiBiT luciferase in the NS1 gene (ZIKV-HiBiT). The HiBiT luciferase sequence with the GS linker was inserted at position 295 aa of NS1. (B) An illustration shows the experimental workflow. Expression of the ZIKV NS1 protein was determined by immunoblotting at 48 hpi of lentiviruses into BHK-21 cells. Luciferase activity in cells and infectious titers in the culture supernatants were determined upon infection of ZIKV-HiBiT at MOI = 0.1 into either parental cells (Mock) or cells expressing ZIKV NS1 (NS1) at the indicated timepoints. (C) NS1 proteins from ZIKV, DENV-2, DENV-4, and JEV were expressed in BHK-21 cells. Luciferase activities in the cells and infectious titers in the culture supernatants were determined at 36 hpt of ZIKV-HiBiT into cells expressing these NS1proteins. Statistical significance was assessed using Student’s t-test (B) or one-way ANOVA with Dunnett’s test (C) and is indicated by asterisks (*) (versus control cells).