*2.6. PDT-AlPc-NE In Vitro*

CT26 or 4T1 cells were incubated in the dark at 37 ◦C with different concentrations of AlPc-NE for 15 min. Then, the cells were washed with PBS and maintained in an incubator for further 15 min with complete RPMI 1640 medium. Next, the cells were irradiated with LED 660 nm for 10 min, with a final energy dose of 25 J/cm<sup>2</sup> . The AlPc-NE concentrations that reduced cell viability by 50% (IC50) and 90% (IC90) were then calculated. In subsequent experiments, the cells were treated with PDT protocols, with their respective AlPc-NE IC<sup>50</sup> and IC90, maintained in an incubator and analyzed for cell death pathways and DAMP exposure.
