Fig 5. Promotion of the initial stages of apoptosis by the activation of our Dual Targeting Intron Constructs.

Clonal Ae. albopictus C6/36 cells transformed with the single or dual targeting antiviral intron constructs indicated were challenged with eeach of the arboviruses shown on the figure. Cells were stained with FITC conjugated annexin V and analyzed the manufacturer’s instructions (Cayman Chemical Company; see Materials and Methods). ΔC/D- or ΔD/C refer to the anti- CHIKV/DENV dual targeting introns possessing the inactive deletion mutation of the trans-splicing domain that is linked to the ΔN Bax 3’ exon. Wt (wild type) refers to a negative control C6/36 cell line that does not express antiviral intron constructs. Six individual clonal cell populations per single or dual virus targeting intron construct are shown. Each number corresponds to the individual clonal cell population tested. Three wells were infected per clonal cell population and analyzed for annexin V-FITC staining. The experiment was performed at three independent times resulting in a total of nine replicates for each dual-targeting intron-ΔN Bax tested.