Figure 6
Miltirone triggers the mitochondrial dysfunction pathway.
(a–b) Exposure of Jurkat, U937 and HL-60 cells to miltirone (0–7.5 μM) for 3 h caused disruption of MMP as evidenced by an increase in the proportion of cells with green fluorescent light (a) and decrease in the proportion of cells with higher red (JC-1 aggregates) / green (JC-1 monomers) ratio of JC-1 fluorescence (b). (c) Cells were treated with 7.5 μM miltirone for 3 h in the presence or absence of 5 mM NAC and the dissipation of MMP was measured. (d–e) Jurkat and U937 cells were treated with miltirone (0–7.5 μM) for 12 h, cytosolic fractions (d) and whole-cell lysates (e) were obtained and subjected to western blot analysis using antibodies against cytochrome c (Cyt c), Bax, Bcl-2 and β-actin; n = 3. The bands were excised from different gels which were run under the same electrophoresis condition. (f) A remarkable Bax / Bcl-2 ratio increasing was demonstrated. Data were presented as Mean ± SD. The differences were significant at *p < 0.05, **p < 0.01 vs. control and # p < 0.05, ## p < 0.01 miltirone vs. co-treatment with miltirone and NAC.
