Cellular ferroptosis inhibition assay

The cell killing capacity of CPMNSs was first evaluated where the well-cultured adherent 4T1 cells were incubated with CPMNSs (25, 50, 100, 125, 150, 175, 200, and 250 μg/ml) for 24 hours, and then, the cell viability was evaluated by CCK-8.

For ferroptosis assay, 4T1 cells were preincubated in 96-well plate for 24 hours and then coincubated with CPMNSs (150 μg ml⁻¹) and numerous types of ferroptosis inhibitors including DFOM (300, 400, and 800 μM), GSH (0.625, 1.25, and 2.5 mM), NAC (3, 4, and 5 mM), Lip-1 (2, 4, and 8 μM), VE (12.5, 25, and 50 μM), and EDTA (200, 400, and 600 μM). After being cultured for another 24 hours, the cell viability was determined by CCK-8.