Pi apoptosis detection kit 1

For cell apoptosis assays, the fluorescein isothiocyanate (FITC)–Annexin V and propidium iodide (PI) Apoptosis Detection kit I (BD Pharmingen) was used as previously described [29 (link)]. For cell cycle analysis, PEL cell pellets were fixed in 70% ethanol, and incubated at 4°C overnight. Cell pellets were re-suspended in 0.5 mL of 0.05 mg/mL PI plus 0.2 mg/mL RNaseA and incubated at 37°C for 30 min. Cell cycle distribution was analyzed on a FACS Calibur 4-color flow cytometer (BD Bioscience).

Cell proliferation was measured by using the WST-1 assays (Roche) according to the manufacturers' instructions. For apoptosis assays, the FITC-Annexin V and propidium iodide (PI) Apoptosis Detection Kit I (BD Pharmingen) was used. Samples were analyzed on a FACS Calibur 4-color flow cytometer (BD Bioscience).

Rat anti-mouse mAbs (Affymetrix eBioscience), which included fluorescein isothiocyanate- (FITC-) labeled anti-CD4 (0.3 μl, No11-0041) as well as phycoerythrin- (PE-) labeled anti-CD8a (0.7 μl, No11-0081), were utilized to stain cells from blood specimens for a 15 min period in dark. After adding hemolysin (250 μl), the cells were subject to further 15 min incubation in the dark and PBS rinsing thrice. CD4⁺/CD8⁺ ratio was determined as the ratio of average fluorescence intensity of CD4⁺ lymphocytes to that of CD8⁺ cells detected using the flow cytometer (Beckman coulter, Navios, USA).
THCA cells undergo certain treatments, including staining using Annexin V-FITC and propidium iodide (PI) Apoptosis Detection Kit I (BD, USA) in line with specific instructions, and cell apoptosis was analyzed through FACS (BD, USA).
Data analysis was completed using Cell Quest Research Software (BD, USA).

Cells were harvested and washed with precooled PBS. Then, the cells were stained with 5 μl of annexin V‐FITC (annexin V‐FITC/propidium iodide (PI) apoptosis detection kit I, BD Biosciences) according to the manufacturer's instructions.