MMP loss, PS externalization, and activation of caspase 3 in 4T1 cells induced by P2Et fraction. (a) 4T1 cells treated with P2Et fraction for 6, 12, 24, or 48 h, stained with JC-1, and analyzed by flow cytometry. Ethanol (negative control) and valinomycin (positive control) (***p < 0.001). (b) PS externalization in 4T1 cells by flow cytometry after treatment for 24 h and 48 h with P2Et fraction, doxorubicin (positive control), or ethanol (negative control). Top panel is a representative dot plot showing the various treatments; bottom is a representative histogram showing percent (%) PS/PI cells after 24 and 48 h of treatment. (c) Caspase 3 activity by enzyme-linked immunosorbent assay (ELISA) after cell treatment with P2Et fraction or doxorubicin (positive control) for 48 h (***p <0.001).