Dcfh da fluorescence probe

The generation of ROS was measured using a DCFH-DA fluorescence probe (Beyotime Institute of Biotechnology, Beijing, China). After being incubated with DCFH-DA (10 µM) for 20 min at 37 °C in a humidified atmosphere in the dark, cells were analyzed with flow cytometry using excitation at 480 nm and emission at 525 nm.

Generation of ROS and SOG were assessed by a DCFH-DA fluorescence probe (Beyotime Institute of Biotechnology, Shanghai, China), and singlet oxygen sensor green reagent (SOSGR; Thermo Fisher Scientific, Inc.) via a fluorescence spectrophotometer (F-4500; Hitachi, Ltd.). SGC-7901 cells were seeded in six-well plates at a density of 2.5×10⁵ cells/well and incubated to adhere securely. Then, the cells were treated with free-AlPcS₄, AlPcS₄ in the presence of 5-FU (20 µM), DOX (0.4 µg/ml), CDDP (5 µM), MMC (0.5 µg/ml), and VCR (0.1 µg/ml) at 1–32 µg/ml for 6 h. The cells were washed twice with PBS and irradiated with laser systems for 5 min. For the detection of ROS, the cells were harvested, incubated with 10 µmol/l DCFH-DA for 20 min at 37°C in complete darkness again, washed with PBS twice, and assessment was conducted using a fluorescence spectrophotometer under an excitation of 488-nm of light. For the detection of SOSGR, the cells were harvested, permeabilized with 0.5% Triton X-100 in PBS for 10 min, centrifuged at 70 × g for 5 min, washed with PBS, mixed with SOSGR, irradiated with a 635-nm laser system for 5 min, and then detected using a fluorescence spectrophotometer under an excitation of 504-nm of light.