Figure 3. APLP2 reduces the sensitivity of TC71 Ewing sarcoma cells to irradiation-mediated apoptosis. (A) TC71 cells were seeded at 5 × 10⁶ cells per 100 mm dish, and upon reaching 40–50% confluence at about 24 h they were transfected with the pCMV-Tag4A vector alone or with pCMV-Tag4A-APLP2. The transfected cells were collected at 48 h and lysed for use in western blots for APLP2 (and for actin, as a control). (B) TC71 cells at 40–50% confluence were transfected with the pCMV-Tag4A empty vector or with pCMV-Tag4A-APLP2 and incubated for 48 h. The cells were then irradiated (0 Gy or 20 Gy), incubated for an additional 24 h, harvested, fixed, stained with propidium iodide, and analyzed for DNA content by flow cytometry. The results in the graph depict the percentage of cells with sub-G₁ DNA content in APLP2-overexpressing cells vs. vector only-transfected cells. Duplicate samples were used and error bars denote the percent confidence interval. The results shown are representative of results from 3 separate experiments.