Annexinv fitc pi double staining kit

Cell death after photodynamic effect was quantified for selected condition using the AnnexinV-FITC/PI double staining kit (Sigma Aldrich). The cell death of LLC cells was investigated 4 h after photodynamic treatment. Briefly, the LLC cells (1 · 10⁵ per well) were cultured overnight in 12-well plates. Then cells were subjected to redaporfin-mediated photodynamic effect (described above), collected by centrifugation and washed twice in Hank’s Balanced Salt Solution (HBBS). The cells were then resuspended in 500 μL binding buffer and stained with annexin V-FITC and PI according to the manufacturer’s instructions (Sigma Aldrich). Stained LLC cells were then examined using Guava® easyCyte™ flow cytometer. Obtained data were analyzed using FlowJo 10.5.3 software (BD Bioscience).

The death of cells was analyzed using the Annexin V-FITC/PI double staining kit (Sigma-Aldrich, USA). H9C2 cells (1 × 10⁶ cells/well) were collected and rinsed twice using cold PBS and then incubated in the Annexin V/PI labeling reagent for 20 min at room temperature in darkness. The cell cycles were detected by flow cytometry (FACSCanto II Analyzer, BD, USA).