![Figure 2. Analysis of caspase activation after treatment with sFCB and TRAIL. 3.5 × 105 KOB cells/mL were incubated for 48 hours with 2.5 μg/mL of sFCB [F], 500 ng/mL of TRAIL [T], simultaneous addition [F + T], or sequential [F→T], or without reagents [C]. (A) Western blot analysis. Using 30 μg of the cell extract and 5 μg of the cytosolic fraction, caspases, and cytochrome c were detected, respectively. (B) Mitochondrial membrane potential (Δψm). After treatment, cells were evaluated using DiOC6(3) by FCM and loss of Δψm (%) is indicated in each panel. (C) Caspase-9 activity. After treatment, cells were evaluated using FITC-LEHD-fluoromethyl ketone by FCM. Shaded and unshaded peaks correspond to treated and untreated cells, respectively. RFI is indicated in each panel. (D) Detection of Bax activation. After treatment, cells were evaluated using a conformation-specific Bax antibody by FCM. Shaded and unshaded peaks correspond to specific and isotype-matched control staining, respectively. RFI is indicated in each panel.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/110/5/10.1182_blood-2007-01-068981/6/zh80180707250002.jpeg?Expires=1724971215&Signature=Ru2f1TgHl0x~4CDt2bDX1fEJnm8V0pTbioKg~3aLH0O8xBDPK31NXdbu33jxC9UsNv0tza8b83o6STSUGNma7wUtv1-95A6XKl2zwPgpSDkNZJz98w1kBuNS-~gtlsl3V0937HblXoChWTHsDqYsl5GcEenmhMIs8NmdvLdBvImvvHIHn40k-8wRDoj1teEXBLu208T-7kvlxeMiOIdgu6KVD197fbJHh~qdvQHmyj56140ZDQ14hT0kn3w7oUsw7-5Em1rXEfUnFSdBOCnEgyiJyS89b2F-DTyxn1MmECeq2LR5o3fgzwzjgxMOdvAWeCsNb80VBQU27kW7GidbbQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Analysis of caspase activation after treatment with sFCB and TRAIL. 3.5 × 105 KOB cells/mL were incubated for 48 hours with 2.5 μg/mL of sFCB [F], 500 ng/mL of TRAIL [T], simultaneous addition [F + T], or sequential [F→T], or without reagents [C]. (A) Western blot analysis. Using 30 μg of the cell extract and 5 μg of the cytosolic fraction, caspases, and cytochrome c were detected, respectively. (B) Mitochondrial membrane potential (Δψm). After treatment, cells were evaluated using DiOC6(3) by FCM and loss of Δψm (%) is indicated in each panel. (C) Caspase-9 activity. After treatment, cells were evaluated using FITC-LEHD-fluoromethyl ketone by FCM. Shaded and unshaded peaks correspond to treated and untreated cells, respectively. RFI is indicated in each panel. (D) Detection of Bax activation. After treatment, cells were evaluated using a conformation-specific Bax antibody by FCM. Shaded and unshaded peaks correspond to specific and isotype-matched control staining, respectively. RFI is indicated in each panel.
