Journal: Scientific reports
Article Title: Toxoplasma GRA16 attenuates Tau hyperphosphorylation and enhances autophagy in thrombin-treated HT-22 hippocampal neuronal cells.
doi: 10.1038/s41598-025-00271-4
Figure Lengend Snippet: Fig. 2. GRA16 upregulated PP2A and PTEN but downregulated GSK3β in thrombin-treated HT-22 cells. (A) GRA16 protein expression in the experimental HT-22 cell groups (control, vector, and GRA16) after thrombin treatment based on Western blot analysis; (B) PP2A-B55 protein expression in the experimental HT-22 cell groups after thrombin treatment based on Western blot analysis and their relative protein expression (n = 3); (C) Western blot image of PTEN and p-PTEN (Ser380) in the experimental HT-22 cell groups after thrombin treatment and their relative protein expression (n = 3); (D) Western blot image of GSK3β and p-GSK3β (Ser389) in the experimental HT-22 cell groups after thrombin treatment and their relative protein expressions (n = 3). †Significant at P < 0.05 between the control and GRA16 in the thrombin-treated experimental HT-22 cell groups. ‡ Significant at P < 0.05 between the vector and GRA16 in thrombin-treated experimental HT-22 cell groups.
Article Snippet: Using western blotting, we Antibody dilution Source Cat. no. Primary antibody Phospho-tau (Ser202, Thr205) (AT8) 1:1,000 Invitrogen MN1020 Phospho-tau (Ser422) 1:1,000 Invitrogen 44-764G Tau 1:1,000 Invitrogen PA5-29610 Phospho-NF-κB p65 (Ser536) 1:1,000 ABclonal PA1294 NF-κB p65 1:1,000 Cell Signaling Technology 8242 HA tag (For GRA16) 1:1,000 Cell Signaling Technology 3724 PP2A-B55 1:1,000 Santa Cruz Biotechnology sc-365,282 Phospho-PTEN (Ser380) 1:1,000 Santa Cruz Biotechnology sc-377,573 PTEN 1:1,000 Santa Cruz Biotechnology sc-7974 Phospho-GSK3β (Ser389) 1:1,000 Proteintech 14850-1-AP GSK3β 1:1,000 Cell Signaling Technology 9315 APOE 1:1,000 Santa Cruz Biotechnology sc-390,925 FOXO3A 1:1,000 Santa Cruz Biotechnology sc-48,348 Caspase 3 1:1,000 Santa Cruz Biotechnology sc-56,053 β-actin 1:1,000 Santa Cruz Biotechnology sc-47,778 Secondary antibody m-IgGK BP-HRP 1:2,000 Santa Cruz Biotechnology sc-516,102 Goat antirabbit IgG (H + L)-HRP 1:4,000 Invitrogen 32,460 Donkey antimouse IgG (H + L)-Alexa Fluor 594 1:2,000 Invitrogen A-21,203 Table 2.
Techniques: Expressing, Control, Plasmid Preparation, Western Blot
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