Figure 6

Effect of 3-NAntC in apoptosis/necrosis profile of the MDA-MB-231 cell line. (a–c) A flow cytometry with Annexin V and Propidium iodide labeling was performed. Cells were treated with the peptide 3-NAntC (0–0.8 µg/mL) or with doxorubicin (0.2 µg/mL) for 24, 48, and 72 h. 3-NAntC induced cell death by apoptosis with a very low amount of necrosis. Data are shown as a mean ± SEM of at least three independent assays. (d) LDH release assay. Cells were treated with the 3-NAntC peptide (0–1.0 µg/mL) for 24, 48, and 72 h and the LDH release was quantified. There is no significant difference in this biomarker, suggesting no involvement of either necrosis or pyroptosis as a primary cell death mechanism. Data are shown as mean ± SEM of at least three independent assays in triplicate. Significant differences between control (0 µg/mL) and treated cells are designated as * p < 0.05, *** p < 0.001, and **** p < 0.0001, according to the two-way ANOVA and Dunnett post hoc test.