Fig. 4.

T-006 promotes LMP7 expression and enhances proteasomal activity through the PKA/Akt/mTOR//p70S6K pathway. Inducible PC12/A53T-α-syn cells were treated with 1 μg/ml Dox for 24 h, and then with T-006 (30 μM) for the indicated time periods; the fold change of expression levels of phosphorylated PKA/total PKA (a), phosphorylated Akt/total Akt (b), phosphorylated p-mTOR/total mTOR (c) and phosphorylated p-p70s6k/total p70s6k (d) were detected by Western blotting analysis. (e) PC12/A53T-α-syn cells were pretreated with or without the PKA inhibitor H-89 (10 μM) for 1 h and then treated with T-006 (30 μM) for 30 min. (f) PC12/A53T-α-syn cells were pretreated with or without the mTOR inhibitor Rap (0.2 μM) for 1 h and then treated with T-006 (30 μM) for 30 min. The fold change of expression levels of phosphorylated p70S6K/total p70S6K were detected by Western blotting. PC12/A53T-α-syn cells were pretreated with or without H-89 (10 μM) for 1 h and then treated with T-006 (30 μM) for 24 h. The protein levels of LMP7 were determined by Western blotting (g). Cellular chymotrypsin-like activity was determined by a commercialized kit (h). Representative Western blotting and data analysis of three independent experiments are shown. Data are represented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001