Figure 3

Nuclear bbtCaspase3-like induced apoptosis independent of its DED. (A) Comparison of domain structures among bfCASP-3/7, human caspase-3, bbtCaspase3-like, and its mutants. (B) BbtCaspase3-like-GFP and C3-2-GFP both localized in nucleus, while C3-1-GFP only containing DED localized ubiquitously in cells. These indicated that bbtCaspase3-like expressed in nucleus independent of its DED. (C) and (D) Caspase-3 and caspase-8 activity of HeLa cells tranfected with different plasmids. (E) The photograph and magnified view of HeLa cells displaying predominantly apoptotic morphology after transient transfection with plasmid encoding bbtCaspase3-like-GFP (20 ×, 63 × objectives). The morphology of cells transfected with GFPN1 vector was as the control. Apoptosis rates were expressed as annexin V positive cells/10,000 gated cells. Data were shown as percent control of the means ± standard deviation of three samples. The negative control was transfected with pCMV vectors and the positive control was treated with camptothecin at 10 μM for 24 h. The third bar represented the sample in the presence of 50 μM z-DEVD-fmk.