Suppressive effect of M₄N on autophagy.

<p><b>A:</b> Effect of M₄N on autophagy in LNCaP cells. The expression of LC3B-I and II was examined by the western blotting in LNCaP cells treated with combination treatments of M₄N with etoposide, rapamycin, or UCN-01 for 18 h in the presence of bafilomycin A₁ (100 nM), an autophagosome degradation inhibitor. Bafilomycin A₁ was added to measure the net activity of autophagy. The concentrations of M₄N, etoposide, rapamycin, and UCN-01 were 80, 20, 20, and 5 μM, respectively. C: Control, E: etoposide, Ra: rapamycin, U: UCN-01. <b>B:</b> The mRNA expression of <i>BNIP3</i> gene, examined by northern blotting, in LNCaP cells treated with M₄N (80 μM) under normoxic or hypoxic condition for 2 or 6 h. Hyp: hypoxic condition. <b>C:</b> The expression of BNIP3 and BNIP3L, examined by the western blotting, in LNCaP cells treated with M₄N (80 μM) under normoxic or hypoxic condition for 6 or 18 h. <b>D:</b> The expression of ATG5, examined by the western blotting, in LNCaP cells treated with M₄N (80 μM) for 18 h. β-Actin was used as a control.