F caspase3

All cell extracts were obtained by lysing cells in ice-cold lysis buffer. The undissolved matter was removed by high-speed centrifugation, and the protein percentage of the supernatant was measured by a BCA protein test kit. Each group of samples was loaded on a 10% SDS-PAGE gel, and the bands were transferred to a PVDF membrane. After being blocked with the sealing solution, the membranes were immunolabeled overnight with the following primary antibodies: HIF-1 (1:1000, Cell Signaling Technology, USA), p53 (1:1000, Cell Signaling Technology, USA), P-gp (1:1000, Cell Signaling Technology, USA), Bax (1:1000, Abcam, USA), F-Caspase3 (1:1000, Cell Signaling Technology, USA), C-Caspase3 (1:1000, Cell Signaling Technology, USA), F-Casase9 (1:1000, Cell Signaling Technology, USA), C-Caspase9 (1:1000, Cell Signaling Technology, USA), F-PARP (1:1000, Abcam, USA), C-PARP (1:1000, Abcam, USA), and β-actin antibodies (1:1000, Santa Cruz, USA). After washing, the protein bands on the PVDF membrane were incubated with suitable auxiliary antibodies (1:2000, ICN Pharmaceuticals), while an ECL kit was used for color development. The bands were analyzed after exposure to a ChemiDoc XRS+ gel imager (Bio-Rad, USA), and the protein content was expressed as the relative value of the corresponding internal reference band.