Flow cytometry software

Flow cytometry was used to determine the apoptosis of glioma cells. Briefly, 1 × 10⁶ DBTRG-Luc and GBM2 cells were seeded in six-well plates and left 24 h in an incubator to resume their exponential growth. Synthetic TRAIL-mRNA, PTEN-mRNA, and PTEN-TRAIL-mRNA were transfected into DBTRG-Luc and GBM2 cells and incubated for 24 h. The cells were then harvested and washed with PBS, and the extent of apoptosis was measured using the Annexin V-FITC Apoptosis Detection Kit (Byotiome, Shanghai,China) and analyzed by flow cytometry software (Beckman, Brea,USA). The upper right part represents apoptotic cells undergoing secondary necrosis at the last stage or dead cells (annexin-V and propidium iodide [PI] double-positive), and the lower right part represents the early-stage apoptotic cell population (annexin-V-positive and PI-negative). Death receptor DR4 and DR5 were also detected by flow cytometry at 12 h and 24 h after TRAIL-mRNA and/or PTEN-mRNA transfected into DBTRG-Luc and GBM2 cells.

Briefly, 1 × 10⁶ DBTRG-Luc cells were seeded in a six-well plate and incubated for 24 h to resume exponential growth. Synthetic TRAIL-mRNA was transfected into DBTRG-Luc cells and incubated for additional 24 h, then the cells were harvested and washed with PBS. The extent of apoptosis was measured through annexin V-FITC apoptosis detection kit (Beyotime, China), and analyzed by flow cytometry software (Beckman, United States). The upper right part represents apoptotic cells undergoing secondary necrosis at the last stage or dead cells (annexin V and phosphatidylinositol [PI] double positive), and the lower right part represents the early-stage apoptotic cell population (annexin V positive and PI negative).