Lsr fortsessa

Apoptosis was analyzed by flow cytometry using the double staining with lipophilic Annexin V and 7-amino actinomycin D (7-AAD) (Abcam, Cambridge, United Kingdom), as described elsewhere (Wang et al., 2017 (link)). Briefly, after treatment, cells were harvested and washed with FACS buffer. Thereafter, cells were resuspended in 100 μl binding buffer (eBioscience) with 2.5 μl Annexin V (eBioscience) and incubated for 20 min at room temperature. 7-AAD (5 μl) was added 5 min before analysis. Early apoptotic cells (Annexin V-positive, 7-AAD-negative), necrotic/late apoptotic cells (double-positive), and living cells (double-negative) were determined by flow cytometry (BD LSR Fortsessa, BD Bioscience, Franklin Lakes, NJ, United States).

Apoptosis was analyzed by using the double staining with the lipophilic dye Annexin V and 7-amino actinomycin D (7-AAD) (Abcam, Cambridge, UK). Briefly, 0.5 × 10⁶ cells were treated with AF for 2 h, harvested by trypsin, washed twice with PBS and resuspended in 100 μl. Thereafter, 2.5 μl Annexin V-FITC and 5 μl 7-AAD were added to the cell suspensions and incubated for 20 min at room temperature in the dark. Early apoptotic cells (Annexin V-positive, 7-AAD-negative), necrotic/late apoptotic cells (double-positive), and living cells (double-negative) were determined by flow cytometry (BD LSR Fortsessa, BD Bioscience, Franklin Lakes, USA).