Figure 5.

RL71 activates CaMKK-AMPK-mTOR signaling pathway. (a) Indicated TNBC cell lines were incubated with various concentrations of RL71 for 24 h or in the presence of RL71 (2 μM) for different time courses. The protein levels of p-AMPK, p-mTOR and p-p70S6K were determined by western blot. β-Actin was used as a loading control. *P<0.05, **P<0.01 versus untreated controls. (b, c) MDA-MB-468 cells were treated with RL71 (2 μM) in the absence or presence of (b) STO-609 (25 μM) and (c) compound C (CC, 10 μM) for 24 h, respectively. The protein levels of p-AMPK and LC3B were analyzed using western blot. **P<0.01. (d) MDA-MB-468 cells were pretreated with STO-609 (25 μM, upper) or CC (10 μM, lower) for 2 h before treatment with various concentrations of RL71 for 24 h. Cell viability was determined by MTT assay. The data are the mean±S.D. of three independent experiments. **P<0.01