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. 2012 Dec 3;123(1):224–235. doi: 10.1172/JCI64516

Figure 1. Specific inhibition of GSK3 reduces BACE1 cleavage of APP.

Figure 1

(A) Swedish mutant APP stable cell line 20E2 was cultured and treated with ARA for 24 hours, and cell lysates subjected to Western blot analysis. Full-length APP and the APP CTFs were detected with C20 antibody. β-Catenin was detected by anti–β-catenin antibody. β-Actin was detected by anti-actin antibody AC-15 as the internal control. (B) Quantification of APP C99 generation in 20E2 cells. ARA treatment significantly increased β-catenin levels in a dose-dependent manner, while APP C99 production decreased with increasing ARA dosage. n = 6; *P < 0.05 and **P < 0.01, ANOVA. Aβ ELISA detection of Aβ40 (C) and Aβ42 (D) in conditioned medium from 20E2 cells treated with ARA for 24 hours. ARA treatment reduced Aβ levels in the conditioned medium in a dose-dependent manner. The values are expressed as mean ± SEM. n = 4; *P < 0.05, ANOVA. (E) γ-Secretase activity in 20E2 cells was inhibited by the pharmacological inhibitor L685,458 (GSI). Co-treatment with specific GSK3 inhibitors ARA and G2 reduced C99. Con, control. (F) Quantification of C99 levels. n = 6; *P < 0.05, ANOVA.