Annexin 5 pe 7 aad apoptosis kit

The plasmids employed in this investigation are listed in Table S2. Annexin V-PE/7-AAD Apoptosis Kit was obtained from Vazyme BioTech Co. LTD. (Nanjing, China). MK2206 and SC79 were achieved from SparkJade (Shandong, China). Rabbit Two-step Kit used for immunohistochemistry staining was obtained from ZSJQ-BIO (Beijing, China). ZEN-BIO provided the goat anti-rabbit IgG (H&L) Alexa Fluor 594 utilized for immunofluorescence (Chengdu, China).

Cells were cultured with epirubicin (2 μM) for 48 h, collected, and divided into two groups. One group was transfected with si-ORM1 and the other group was transfected with control siRNA, followed by incubation for 48 h. The annexin V-PE/7-AAD apoptosis kit (Vazyme, Nanjing, China) was used to analyze the cells. The cultured cells were collected into groups, digested with trypsin, and centrifuged. For washing, the supernatant was removed, PBS was added, the cells were resuspended, and the procedure was repeated 3 times. Next, 250 μL of binding buffer was added to the resuspended cells. The cell suspension (100 μL) was mixed with 5 μL Annexin V-PE and 10 μL 7-AAD solution, and incubated for 15 min in the dark. The stained cells were immediately analyzed by flow cytometry. Finally, flowjo software was used for data analysis. The mortality rate in the fourth quadrant was analyzed for each group and compared.