Fig. 3

Mirodenafil downregulates GSK-3β signaling, resulting in reduced tau phosphorylation, decreased Aβ production by inhibiting amyloidogenesis and activating the autophagosomal pathway. A, B Western blot analysis for A Aβ, B phosphorylated tau, and total tau in mouse hippocampal homogenates of the APP-C105 mice (n=4). C Western blot analysis for phosphorylated tau at S199/202 and at S396 in SH-SY5Y cells after co-treatment with okadaic acid and mirodenafil (n=4). D Western blot analysis for phosphorylated GSK-3β and phosphorylated Akt in SH-SY5Y cells after co-treatment with Aβ₄₂, mirodenafil, the PI3K inhibitor LY-294002 (n=4). E Western blot analysis and F real-time qRT-PCR results for APP and BACE1 expression after co-treatment with Aβ₄₂ and mirodenafil in HT-22 cells (n ≥ 5). G Western blot analysis for phosphorylated AMPK and Aβ after co-treatment with Aβ₄₂ and mirodenafil in SH-SY5Y cells (n=4). H Western blot analysis for the autophagy markers p62 and LC3B after co-treatment with Aβ₄₂, mirodenafil, and autophagy inhibitor 3-MA (n=4). Data are presented as the mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. All statistical comparisons were performed relative to Aβ₄₂-treated cells. Statistical significance was assessed by one-way ANOVA followed by Bonferroni’s post hoc test for multiple comparisons