Trehalose induces autophagy and protects Me21 cells from cisplatin-induced apoptosis.

<p>(<b>A</b>) Me21 melanoma cells were treated with 100 mM trehalose (Tre) for 22 hours, then exposed to 200 nM bafilomycin A1 (Baf) for further 2 hours. Western blot of LC3 and LC3-II densitometric analysis of a typical experiment out of 3 are reported; in bafilomycin-treated cells half protein has been loaded. (<b>B, C</b>) Cells were treated with cisplatin for 24 h, with or without trehalose, and with or without 3-MA; (<b>B</b>) total cell numbers are presented as percentage over control cells; the columns also show the relative number of adhering cells (white) and floating dead cells (black); S.E. values and statistical analysis refer to detached cells; (<b>C</b>) caspase-3/-7 activity; data are reported as means ± S.E. of 4 to 5 experiments. *<i>P</i><0.001,**<i>P</i><0.01, and ***<i>P</i><0.05. (<b>D</b>) Control cells were treated with 100 mM trehalose for 24 h, with or without the autophagy inhibitor, 3-methyladenine (3-MA); Western blot of LC3 and densitometric analysis of a typical experiment out of 5 are reported. (<b>E</b>) Cells were treated for 24 hours with cisplatin with or without 100 mM trehalose; cisplatin-treated samples (single or combined treatment) refer to adhering cells. Western blot of LC3 and LC3-II densitometric analysis of a typical experiment out of 4 are reported. Densitometric analyses were normalized to Ponceau S staining.