FIGURE 3.

The mitochondrial pathway is induced by archazolid. A, SKBR3 cells were left untreated (dotted line) or incubated with 1 nm (solid line) and 10 nm (dashed line) archazolid for 24 h, and mitochondrial potential was assessed by JC-1 staining via flow cytometry. An increase in green fluorescence indicates the loss of membrane potential. B, the activation of Bax was measured by flow cytometry (anti-Bax 6A7) after 48 h of treatment. C, the release of cytochrome c from the mitochondria was measured via flow cytometry after 48 h. D, activation of caspase-9 was assessed by cleavage of LEHD-AFC via fluorometry after 24 and 48 h of treatment. E, mitochondrial membrane potential in rat liver mitochondrial suspensions was assessed by rhodamine-123 fluorescence. Increasing amounts of archazolid and concanamycin (conc) were tested; calcium served as positive control. F, ATP productivity was analyzed in isolated rat liver mitochondria after treatment with different concentrations of archazolid or concanamycin. Oligomycin and potassium cyanide (KCN) served as background controls. *, p ≤ 0.05, ANOVA/Dunnett.