Fig. 3: Trehalose and rapamycin induce autophagy in LNCaP cells, but do not induce autophagy in DU145 cells.

a LNCaP cells were treated with 20, 50,100 mM trehalose or with 10, 50, 100 nM rapamycin for 48 h. MTT assays were performed. Data are mean ± SD of six independent biological samples (n = 6). Each experiment was repeated three times. Statistical analysis of data was performed by Dunnet test (*p < 0.05 vs. control). b LNCaP cells were treated for 24, 48 or 72 h with 100 mM trehalose or 100 nM rapamycin and WB analysis of LC3 was performed. Detection of autophagy was analysed by quantification of LC3-II/LC3-I ratio. Relative optical density was determined by ImageJ software. Experiments were performed independently three times and a representative blot is shown. c LNCaP cells were treated for 24, 48 or 72 h with 100 mM trehalose or 100 nM rapamycin and WB analysis of p62 was performed. Tubulin was used as loading control. Bar graph represents mean optical density ± SD of p62 levels. The experiments were performed independently three times. Statistical analysis was performed by one-way ANOVA with Bonferroni post-test (*p < 0.05). d DU145 cells were treated with 20, 50,100 mM trehalose or with 10, 50, 100 nM rapamycin for 48 h. MTT assays were performed. Data are mean ± SD of six independent biological samples (n = 6). Each experiment was repeated three times. Statistical analysis of data was performed by Dunnet test (*p < 0.05 vs. control). e DU145 cells were treated for 48 or 72 h with 100 mM trehalose or 100 nM rapamycin. WB analysis of LC3 was performed. Protein extract of PC3 cells treated with 100 mM trehalose for 48 h was utilized as a positive control, tubulin was used as loading control. f DU145 cells were treated for 48 or 72 h with 100 mM trehalose or 100 nM rapamycin. WB analysis of p62 was performed. Tubulin was used as loading control. Bar graph represents mean optical density ± SD of p62 levels. The experiments were performed independently three times. Statistical analysis was performed by one-way ANOVA with Bonferroni post-test.