Subsequently, Annexin V–FITC/PI double-staining was performed using flow cytometry to evaluate the effect of the apoptosis induced by daphnegiravone D. The results showed that AV+PI− and AV+PI+ cells increased dramatically in a dose-dependent manner in both hepatoma cell lines after treatment with daphnegiravone D for 48 h (Fig. 4A and B), demonstrating that apoptosis was induced. The gene discussed is ANXA5; the disease is hepatocellular carcinoma.