Annexin 5 apc 7 aad apoptosis detection kit

For anoikis analysis, OS cells were cultured in suspension for 48 h. Cells were then trypsinized and stained with Annexin V- APC/7AAD and analyzed by flow cytometry using an Annexin V- APC/7AAD Apoptosis Detection kit (BD Biosciences) according to the manufacturer’s instructions. Data were collected on a BD FACSCan and analyzed using the FlowJo software.

For apoptosis detection, the cells were harvested and stained using an Annexin V-APC/7-AAD apoptosis detection kit (BD Biosciences, San Jose, CA, United States) following the manufacturer’s protocol. FACS analysis was performed by flow cytometry using a flow cytometer (BD FACSCanto II, BD Biosciences, CA, United States) and analyzed using FlowJo (TreeStar) software.

Cell apoptosis was analyzed by flow cytometry. The Annexin V-APC/7-AAD Apoptosis Detection Kit (BD Biosciences) was used in accordance with the manufacturer’s instructions. Briefly, HCC cells receiving different treatments (1 × 10⁵) were harvested, washed twice with cold PBS, and resuspended in a 1× binding buffer. Next, the cells were incubated with 5 μl of Annexin-APC and 5 μl of 7-AAD for 15 min at room temperature in the dark. The cells were immediately analyzed using a FACSCalibur flow cytometer (Becton Dickinson). All experiments were performed in triplicate. Apoptosis in tumor tissues was analyzed by a TUNEL assay, and the apoptotic cells were determined using an in situ apoptosis detection kit (KEYGEN, China) according to the manufacturer’s instructions. The frequency of apoptotic cells in a section of tumor sample was obtained by calculating the mean number of TUNEL-positive brown nuclei among the hepatocytes in five randomly selected fields viewed under a microscope at ×100 magnification.

Cell apoptosis was analyzed by annexin V-APC/7-AAD staining as previously described [58 (link)]. In brief, the serum deprivation medium was used for inducing apoptosis [59 (link)]. Cells of each group seeded on sterile cover glasses were placed in 6-well plates for culture in DMEM medium supplemented with vehicle (PBS), n-EMVs or OGD-EMVs for 24 h. The apoptosis assay of atrocytes was conducted with using an Annexin V-APC/7-AAD apoptosis detection kit (BD Biosciences). Briefly, cells were washed with PBS, resuspended with 100 μL 1 × annexin-binding buffer, incubated with 5 μL APC-conjugated Annexin V and 5 μL 7-Amino-actinomycin (7-AAD) for 15 min in the dark, then analyzed by flow cytometry. Cells stained with both Annexin V-APC and 7-AAD were considered to be late apoptotic astrocytes, and the cells stained only with Annexin V-APC were considered to be early apoptotic astrocytes. The experiment was repeated three times, and three plates per experiment were analyzed in each group.

Cell apoptosis was examined using an Annexin V-APC/7-AAD Apoptosis Detection Kit (BD Biosciences, San Jose, CA, USA) and analyzed by flow cytometry using the BD FACSAria instrument.

Siha-NC (negative control) and Siha-KD (knockdown) were plated in six-well plates, respectively. Medium with and without cisplatin was added to the wells for another 24 h after cell attachment. Then cells were collected for flow cytometry to detect apoptosis. Cell apoptosis was determined by Annexin V-APC/7AAD apoptosis detection kit (BD Biosciences) following the manufacturer’s product instruction manual.