Figure 4.

LSD1 regulates autophagy through the mTOR signaling pathway. (A) The HO8910-shLSD1 cells were treated with different doses of Dox for 48 h. Phosphorylated p70S6K (p-p70S6K; Thr389) and total p70S6K were probed by western blotting. (B) HO8910 cells were treated with 100 μM TCP for 24 h, and phosphorylated p70S6K (p-p70S6K; Thr389) and total p70S6K were probed by western blotting. (C) HO8910-shLSD1 cells were incubated in medium without serum for 16 h or 50 nM rapamycin for 12 h followed by treatment with or without 100 ng/ml Dox for 48 h and the proteins of interest were detected by western blotting. *P<0.05. (D) The HO8910-shLSD1 cells were treated with 10 μΜ mTOR specific activator MHY1485 for the indicated time-points. The expression of p-p70S6K and p70S6K was detected via western blot analysis. (E) The HO8910-shLSD1 cells were pretreated with 10 μM MHY1485 for 1 h prior to the addition of Dox and the proteins of interest were detected via western blotting. *P<0.05. (F) The HO8910 cells were pretreated with 10 μM MHY1485 for 1 h prior to the addition of TCP and the proteins of interest were detected via western blotting. *P<0.05. α-tubulin, loading control. Data shown are representative of three independent experiments.