Beckman gallios flow cytometry

Apoptosis was analyzed as mentioned previously [18 (link)]. T-ALL cells (6T-CEM and J-gamma1 cell lines) were infected with lentivirus for 24 hr in the presence of 10 μg/ml polyurethane (Sigma–Aldrich). Stable cell lines were selected with 10 μg/ml of puromycin (Invitrogen; Thermo Fisher Scientific, Inc.). Leukemia cells obtained after 5 days of screening were washed with precooled 1x PBS and suspended in a 1x binding buffer and then treated using the FITC-Annexin V Apoptosis Kit (item no.: 556420; BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's instructions. Flow cytometry (Beckman Gallios™ Flow Cytometry; Beckman) was used to analyze the apoptosis.

Follow the manufacturer’s instructions. 1 × 106 cells were collected, washed with pre-cooled 1xPBS, and then resuspended in 100 µl of 1 × binding buffer. Subsequently, fluorescein isothiocyanate (FITC)-AnnexinV (5 µl) and PI (5 µl) were sequentially added (cat. No. 556420; BD Biosciences, Franklin Lakes, NJ, USA). The mixture was incubated away from light for 15 min with an additional volume of 400 µl of 1 × binding buffer. Apoptosis analysis was performed using Beckman Gallios^™ flow cytometry (Beckman, Krefeld, Germany) following standard procedures, and the results were analyzed using FlowJo_V10.