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Figure 4 Newly synthesized HDAC inhibitors activate the intrinsic cell death pathway. The proportion of early apoptotic and late apoptotic MDA-MB-231 cells was measured using bivariate Annexin V/7AAD analysis by flow cytometry after the cells underwent treatment with tested compounds at IC50 concentration for (A) 24 h, (B) 48 h, and (C) 72 h. (D) Dissipation of mitochondrial membrane potential was assessed by flow cytometry using JC-1 dye (compared to the untreated control). Significant differences between treatments were determined by t-test: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 (compared to the untreated control).