Annexin 5 fitc kit

U251 and U343 cells were reverse transfected (siRNA control and SERBP1) and harvested 72 h later using trypsin. 2.5 × 10⁵ cells were prepared following an Annexin V-FITC kit (catalog #: A13199, V13246 Thermo Fisher). Annexin staining was subsequently measured by flow cytometry. All experiments were performed with biological and technical triplicates.

Cell apoptosis was tested by the Annexin V-FITC kit (Thermo Fisher Scientific, Inc.). First, SW480 and HT-29 cells were collected, resuspended in cold PBS, centrifugated at 1000 rpm in room temperature for 5-10 min. Then, cells were incubated with 5 μL AnnexinV-Alexa Fluor 647 for 15 min and then co-incubated with 5 μL PI before detection. Finally, the apoptosis rate was analyzed via flow cytometer (Beckman Coulter, CA, USA).

Cells were digested with trypsin and then washed with PBS. Annexin V-FITC Apoptosis Detection Kit (Beyotime) was applied to stain apoptotic cells following the manufacturer's instructions. The apoptotic cells were dual-stained with PI and AnnexinV-FITC, using Annexin V/FITC kit (Thermo Scientific, Shanghai, China). The analysis was carried out via BDTM LSRІІ flow cytometer (BD Biosciences). Afterward, data was measured with the Cell Quest (BD Bioscience, San Jose, CA, USA) software.

Cells were digested with trypsin and then washed with phosphate-buffered saline (PBS). An Annexin V-FITC Apoptosis Detection Kit (Beyotime) was applied to practice cell apoptosis in line with the manufacturer's instructions. The apoptotic cells were dual-stained with propidium iodide (PI) and Annexin V-FITC using an Annexin V-FITC Kit (Thermo Scientific, Shanghai, China). Analysis was carried out via BDTM LSRII flow cytometer (BD Biosciences). Afterward, the data were measured with the Cell Quest software (BD Bioscience, San Jose, CA, USA).

1 × 10⁶ cells were plated in 25 cm² angled-flasks, except VCaP cells (1.5 × 10⁶ cells). 48 and 72 h after treatments, cells were trypsinized. We performed the assay with Cell cycle test kit (BD Biosciences, California USA) according to manufacturer’s instructions. Nocodazole (Sigma, St. Louis, MO, USA) at 1 μM was used to arrest the cell cycle in G2 phase. We used half of the cell population fated to apoptosis assays. In this case, cells were processed using Annexin V- FITC Kit (Thermo Fisher Scientific, V13242) according to manufacturer’s instructions. 2-amino-N-quinoline-8-yl-benzenesulfonamide (QBS) (SIGMA) was selected as an apoptosis inducer. Attune (Applied Biosystems, Life Technologies, Carlsbad, CA) was used for performing cell cycle and apoptosis assays.

Cells in 2.3 were collected and performed according to the Annexin V-FITC kit (Thermo Fisher Scientific, Carlsbad, CA, USA). The apoptosis of B16F10 cells was analyzed by flow cytometry (Ex = 488 nm, FL1 Em = 525 ± 20 nm, FL2 Em = 585 ± 21 nm) and FlowJo software.

Human AML U937, AML HL-60, AML THP-1, CML K562, and ALL Jurkat cells from BCRC (Hsinchu, Taiwan) were grown in RPMI-1640 medium supplemented with 10% FCS, 2 mM L-glutamine, 1% sodium pyruvate, and 1% streptomycin/penicillin. Cells were maintained in a humidified incubator with a 95% air/5% CO₂ atmosphere at 37 °C. Cell viability was detected using a MTT assay [66 (link)]. Cell apoptosis was detected by staining cells with annexin V-FITC kit (Thermo Fisher Scientific) followed by flow cytometric analysis. The ABT-199-resistant U937 cells were prepared according to the procedure described in our previous studies [67 (link)]. Cell culture supplements and media were the products of GIBCO/Life Technologies Inc. (Carlsbad, CA, USA). The morphologies of U937, HL-60, THP-1, K562, Jurkat, and ABT-199-resistant U937 cells are shown in Supplementary Figure S4.