Figure 3. Assessment of apoptosis by BxPC3 cells were treated with F3 or F3.CE and CPT-11.

BXPC3 cells were treated with or without F3.CE cells at the indicated doses for 48 hrs. A. Cell viability was measured with propidium iodine (PI). Significantly less percentage of BXPC3 cells were viable when treated with 1 μM CPT-11 and F3.CE, compared to F3. B., C. annexin V-based apoptosis assay was performed as described in Materials and Methods. BXPC3 cells treated with CPT-11 and F3.CE showed significantly increased ratio of apoptotic cells. D. Bystander effect of the CE produced by the F3.CE cells was confirmed using a co-culture system of F3 or F3.CE cells and BXPC3 pancreas adenocarcinoma cells. BXPC3 cells with F3 or F3.CE cells were seeded in 96-well plates (total 1 × 10⁴ cells per well, BXPC3 cells: F3 or F3.CE cells = 100:0, 75:25, 50:50, 25:75, or 0:100). After 48 hrs in co-culture, cells were treated with 1.0 μM/mL CPT-11 for 48 hrs and cell survival was determined (each group, n = 3) (p< 0.05).