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CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the <t>Baf-A1,</t> CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.
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CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the <t>Baf-A1,</t> CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.
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CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the <t>Baf-A1,</t> CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.
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CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the <t>Baf-A1,</t> CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.
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CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the <t>Baf-A1,</t> CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.
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Image Search Results


CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the Baf-A1, CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.

Journal: Frontiers in Aging Neuroscience

Article Title: Clemastine alleviates Parkinson’s disease pathology by activating the AMPK/mTOR pathway and enhancing autophagic flux

doi: 10.3389/fnagi.2026.1915475

Figure Lengend Snippet: CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the Baf-A1, CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.

Article Snippet: The CTRL group was treated with normal saline (NS; volume equal to that of MPP + ), while the PD model group was treated with MPP + at a final concentration of 1 mM for 24 h. Cells in both CTRL and MPP + groups were subsequently treated with CLE, Baf-A1 (Selleck), or dorsomorphin (DOR; Selleck) according to the experimental design.

Techniques: Phospho-proteomics, Western Blot, Analysis, Control, Expressing, Comparison