The silencing of CASP8AP2 / FLASH activates markers of apoptosis. The effect of silencing CASP8AP2/FLASH (two different siRNAs) for (A) 48 and (B) 72-hours on the levels of (i) activated Caspase 8, (ii) Caspase 3/7 and (iii) cell viability. The viability of cells transfected in parallel is shown for comparison and a siRNA corresponding to PLK1 was included as a positive control. Five independent transfections of each siRNA were performed. The statistical comparison (t-test, unequal) of siNeg transfected cells to CASP8AP2/FLASH, NUP62 or PLK1 silenced cells is indicated by *** = p ≤0.001, ** = p ≤0.01, and *. = p ≤0.05. Caspase 8 levels were measured using an Ac-LETD-pNA Caspase-8 substrate (Caspase-Glo 8 Assay, Promega), Caspase 3/7 levels were measured using a DEVD peptide substrate (Caspase-Glo 3/7 Assay, Promega), and cell viability was measured using Cell Titer Blue Reagent, Promega).