Journal: International Journal of Molecular Sciences

Article Title: Increase in BACE1 Phosphorylation in the Rat Hippocampus and Phytosphingosine in Plasma After Chronic Administration of Scopolamine

doi: 10.3390/ijms27052241

Figure Lengend Snippet: Expression of BACE1 and P-BACE1-T252 in the hippocampus. BACE1 and β-actin bands obtained by Western blotting in the control and Tx groups ( A ); P-BACE1-T252 and β-actin bands obtained by Western blotting in the control and Tx groups ( B ); quantitation of the BACE1 bands shown in A ( C ); quantitation of the P-BACE1-T252 bands shown in B ( D ); levels of P-BACE1-T252 obtained by ELISA ( E ). * p < 0.05; ** p < 0.01; *** p < 0.001. n = 8 (control group); n = 9 (Tx group).

Article Snippet: Electrophoresis, transfer, and blocking were performed as described above; the membrane was then incubated with a primary antibody against phospho-BACE1 (T252) (Affinity Biosciences, Cincinnati, OH, USA, AF3650, 1:1000 dilution in blocking solution) with constant agitation for 12 h. After five washes with PBST (10 min each), the membrane was incubated with an HRP-conjugated goat anti-mouse IgG (H + L) secondary antibody (Novus Biologicals HRP NB730-H, 1:1000 dilution in PBST) with constant agitation at room temperature.

Techniques: Expressing, Western Blot, Control, Quantitation Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Increase in BACE1 Phosphorylation in the Rat Hippocampus and Phytosphingosine in Plasma After Chronic Administration of Scopolamine

doi: 10.3390/ijms27052241

Figure Lengend Snippet: Expression of BACE1 and P-BACE1-T252 in the prefrontal cortex. BACE1 and β-actin bands obtained by Western blotting in the control and Tx groups ( A ); P-BACE1-T252 and β-actin bands obtained by Western blotting in the control and Tx groups ( B ); quantitation of the BACE1 bands shown in A ( C ); quantitation of the P-BACE1-T252 bands shown in B ( D ); levels of P-BACE1-T252 obtained by ELISA ( E ). * p < 0.05; *** p < 0.001; ns = not significant. n = 8 (control group); n = 9 (Tx group).

Article Snippet: Electrophoresis, transfer, and blocking were performed as described above; the membrane was then incubated with a primary antibody against phospho-BACE1 (T252) (Affinity Biosciences, Cincinnati, OH, USA, AF3650, 1:1000 dilution in blocking solution) with constant agitation for 12 h. After five washes with PBST (10 min each), the membrane was incubated with an HRP-conjugated goat anti-mouse IgG (H + L) secondary antibody (Novus Biologicals HRP NB730-H, 1:1000 dilution in PBST) with constant agitation at room temperature.

Techniques: Expressing, Western Blot, Control, Quantitation Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Increase in BACE1 Phosphorylation in the Rat Hippocampus and Phytosphingosine in Plasma After Chronic Administration of Scopolamine

doi: 10.3390/ijms27052241

Figure Lengend Snippet: BACE1 activity. BACE1 activity in the hippocampus of rats treated with scopolamine (Tx group) was significantly greater than that in the control group. ** p < 0.05; n = 8.

Article Snippet: Electrophoresis, transfer, and blocking were performed as described above; the membrane was then incubated with a primary antibody against phospho-BACE1 (T252) (Affinity Biosciences, Cincinnati, OH, USA, AF3650, 1:1000 dilution in blocking solution) with constant agitation for 12 h. After five washes with PBST (10 min each), the membrane was incubated with an HRP-conjugated goat anti-mouse IgG (H + L) secondary antibody (Novus Biologicals HRP NB730-H, 1:1000 dilution in PBST) with constant agitation at room temperature.

Techniques: Activity Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Increase in BACE1 Phosphorylation in the Rat Hippocampus and Phytosphingosine in Plasma After Chronic Administration of Scopolamine

doi: 10.3390/ijms27052241

Figure Lengend Snippet: Modifications in protein expression, activity and metabolite levels in the scopolamine model. Scopolamine administration blocks M1 receptors, promotes amyloidogenic APP processing through increased BACE1/P-BACE1 and β-amyloid production, induces mitochondrial dysfunction with metabolomic alterations (↑ phytosphingosine, ↓ acetylcarnitine), and suppresses the cAMP/PKA/CREB pathway, leading to reduced BDNF expression and neuroprotection.

Article Snippet: Electrophoresis, transfer, and blocking were performed as described above; the membrane was then incubated with a primary antibody against phospho-BACE1 (T252) (Affinity Biosciences, Cincinnati, OH, USA, AF3650, 1:1000 dilution in blocking solution) with constant agitation for 12 h. After five washes with PBST (10 min each), the membrane was incubated with an HRP-conjugated goat anti-mouse IgG (H + L) secondary antibody (Novus Biologicals HRP NB730-H, 1:1000 dilution in PBST) with constant agitation at room temperature.

Techniques: Expressing, Activity Assay, Metabolomic