Annexin 5 apc propidium iodide apoptosis detection kit

The SNU620, MKN45, MKN28, KATO III, and AGS gastric cell lines were obtained from the Korean Cell Line Bank (Seoul, Korea) and maintained in RPMI 1640 supplemented with 10% fetal bovine serum. The cells were cultured in 100% humidity and 5% CO₂ at 37 °C. The c-MET inhibitor drug (tepotinib) was purchased from Selleck Chemicals (Houston, TX, USA). The annexin V-APC/propidium iodide (PI) apoptosis detection kit (Thermo Fisher Scientific, Rockford, IL, USA) was used in the present study.

HAP1, HAP1 RNF43 KO, HAP1 PWWP2B KO, and GC (SNU620 cells: high-RNF43, low-PWWP2B; Kato III cells: middle-RNF43 and middle-PWWPB; MKN28: low-RNF43 and high-PWWP2B; and MKN45 cells: low-RNF43 and low-PWWP2B) cells seeded onto 6-well plates at a density 5 × 10⁴ cells per mL were treated with the respective IC50 values of docetaxel trihydrate, pelitinib, and uprosertib (Table 2). Cell death was determined using the Annexin V-APC/Propidium Iodide (PI) Apoptosis Detection Kit (Thermo Fisher Scientific, Rockford, IL, USA) on a CytoFLEX flow cytometer (Beckman Coulter, Brea, CA, USA). The percentages of intact and apoptotic cells were calculated using CytExpert software (Beckman Coulter).