Fig. 5.

The antitumor effect of ES@Cu(Ⅱ)-MOF-induced cuproptosis in vitro.

A-D) Relative viabilities of 4T1, MDA-MB-231, MDA-MB-468, and BEND3 cells following 24 h of incubation with a gradient concentration of ES@Cu(Ⅱ)-MOF NPs (n = 3).

E) Morphological injury of 4T1 cells after different treatments. Scale bar: 50 μm.

F) CLSM images of 4T1 cells stained with calcein-AM (green, live cells) and PI (red, dead cells) after different treatments. Scale bar: 50 μm.

G) Flow cytometric analysis of the degree of necrosis in 4T1 cells following various treatments.

H) Quantification of Annexin V-FITC⁺/PI⁺ 4T1 cells after the indicated treatment (n = 3).

I) Colony formation ability of 4T1 cells after different treatments.

J) Quantification of the number of colonies formed by 4T1 cells after the indicated treatment (n = 3).

4T1 cells treated with CuCl₂ + ES were used as the positive control [3]. The data are presented as the means ± SDs; ∗∗p < 0.01; ∗∗∗p < 0.001.