Annexin 5 apc 7 aad kit

The apoptosis rate was detected using the Annexin V-APC/7-AAD kit (BD, USA). Cells were exposed to 6 MV-X ray at doses of 0 and 4 Gy, and all cells in each group were collected 48 h after irradiation. Then, the cells were labelled with 5 μL Annexin V-APC and 5 μL 7-AAD according to the manufacturer’s instructions. Apoptotic cells were analyzed by the flow cytometry system (BD FACSCalibur, USA) within 1 h.

HepG2 cells (5 × 10⁵cells/well) were seeded in 6-well plates with 1640 medium. After incubated overnight at 37 °C and in an atmosphere of 5% CO₂, the medium was removed, the cells were washed twice with PBS, and the following treatments were applied: blank group, only 1640 medium was added; Curcumin group, 1640 medium containing 10 μg/mL curcumin was added; GAMCLCL group, 1640 medium containing GAMCLCL (equivalent to containing 10 μg/mL curcumin) was added. After incubation for 24 hours, the medium was removed, and the cells were carefully washed twice with PBS. After digestion with 0.25% trypsin without EDTA, the cells were centrifuged for 5 min at 1200 rpm and harvested. The AnnexinV-APC/7-AAD kit (KGA1026, Nanjing, China) was used for the flow cytometry (BD Biosciences, USA) evaluation of apoptosis.