Effect of combination treatments on cell death-related cellular functions.

<p><b>A-B:</b> Analysis of caspase-dependent cell death mechanisms induced by combination treatments of M₄N with Etoposide, Rapamycin, or UCN-01. <b>Aa & Ab:</b> Cleavage of caspase-3, -4, -7, and -9 along with poly-ADP ribose polymerase (PARP) in LNCaP cells treated by the combination treatments for 17 h. β-Actin was used as a control. <b>B:</b> Caspase-7 enzymatic activity in LNCaP cells treated by the combination treatments for 13 h. Data are presented as mean±SD in triplicate. <b>A & B:</b> C: control, E: Etoposide (10 μM), Ra: Rapamycin (10 μM), Ra30μM: Rapamycin (30 μM), U: UCN-01 (2 μM), M4N: M₄N (80 μM), and Casp: Caspase. <b>C:</b> Effect of combination treatments on mitochondrial membrane potential (ΔΨ_m). LNCaP cells were treated with M₄N (80 μM) in combination with Etoposide (10 μM), Rapamycin (10 μM), or UCN-01 (2 μM) for 4 h. The ΔΨ_m was measured using JC-1 dye. The image shows the ratios obtained by dividing the intensity at 568 nm-excitation light (J-aggregates) by that at 488 nm-excitation light (J-aggregates + monomer). The color bar is shown on the right side of the figure (H: high, L: low ratio).