(a) Cells were co-transfected with poly(IC) and C or C-IU (I). Dead cells (supernatant) were counted after 12–24h (n=3). (b) HeLa cells were co-transfected with poly(IC) and C or C-IU, or treated with CPT ± C or C-IU. Immunoblotting was used to detect apoptosis by analyzing PARP cleavage (cPARP). Uncleaved PARP is also shown (PARP). IRF3 activation was analyzed (IRF3 S396-P), and actin was a loading control. (c) HeLa cells were mock transfected (LF-2000), co-transfected with poly(IC) and C or C-IU, or treated with CPT. Cells were labeled after 24h with Annexin V FITC and PI, and analyzed by FACS. The percentage of living (L; Annexin V−, PI−), early apoptotic (EA; Annexin V+, PI−) and late apoptotic (LA; Annexin V+, PI+) cells are given. (d) The proportion of apoptotic (EA+LA) cells observed in samples described in (c) were quantified by FACS, and summarized graphically (n≥3). All error bars are mean ± s.d.