Fig. 7. Effect of amitriptyline on Beclin 1 acetylation and Rubicon binding.

a Differentiated SH-SY5Y cells were pretreated with 10 μM LY294002 for 1 h, followed by 20 μM amitriptyline for 24 h. p300 protein levels were analyzed by western blotting, and quantified by densitometric analysis (n = 3). b Cells were treated with 20 μM amitriptyline for 24 h, and analyzed protein interaction by immunoprecipitation with anti-Beclin 1 and immunoblotting with anti-Beclin 1, anti-acetylated-lysine, and anti-Rubicon, and quantified by densitometric analysis (n = 5). c Schematic diagram representing amitriptyline-mediated inhibition of autophagosome maturation and related signaling pathway. *P < 0.05, **P < 0.01. AMI amitriptyline, LY LY294002, IP immunoprecipitation, IB immunoblotting.