Tunel fluorescein assay kit

DNA fragmentation analysis was carried out by using a TUNEL fluorescein assay kit (Roche Basel, Switzerland). Tissue slides were incubated with TUNEL reaction mixture (terminal deoxynucleotidyl transferase and nucleotide mixture) for 1 h at 37°C in a humidified and dark atmosphere. Following three washes with PBS, the samples were analyzed using a Meta confocal laser scanning microscope (Carl Zeiss LSM 800).

DNA fragmentation analysis was carried out by using a TUNEL fluorescein assay kit (Roche, Basel, Switzerland). PASMCs were seeded onto 24-well plates with a circle microscope cover glass (15 mm) at a concentration of 3 × 10⁵ cells per well. After treatment, PASMCs were washed three times with PBS, fixed with 4% paraformaldehyde for 1 h, and permeabilized with 0.1% Triton X-100 for 5 min. The cells were then incubated with TUNEL reaction mixture (terminal deoxynucleotidyl transferase and nucleotide mixture) for 1 h at 37 °C in a humidified and dark atmosphere. After a rinse with PBS, the samples were analyzed under a Carl Zeiss LSM 800 Meta confocal laser scanning microscope (Carl Zeiss).