Effector caspase activation confirms the induction of apoptosis by the activation of Dual Targeting Intron Constructs.

Clonal Ae. albopictus C6/36 cells transformed with the single or anti-CHIKV/DENV dual targeting antiviral intron constructs indicated were challenged with either of the four DENV serotypes or CHIKV vaccine strain 181/25 (MOI 0.01). 1x106 cells were assessed for effector caspase activity, per the manufacturer’s instructions (see Materials and Methods). ΔC/D-ΔN Bax or Δ/C-ΔN Bax 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. Numbered lanes indicate clonal cell populations expressing single or dual virus targeting intron constructs. Three wells were infected per clonal cell population and analyzed for “effector caspase” activity. The experiment was performed at three independent times resulting in a total of nine replicates for each dual-targeting intron-ΔN Bax tested.