Journal: The Journal of Biological Chemistry
Article Title: Combined treatment with the phenolics (−)-epigallocatechin-3-gallate and ferulic acid improves cognition and reduces Alzheimer-like pathology in mice
doi: 10.1074/jbc.RA118.004280
Figure Lengend Snippet: Combination therapy with EGCG plus FA dampens neuroinflammation and oxidative stress. Data were obtained from APP/PS1 mice that received vehicle ( APP/PS1-V , n = 8), EGCG ( APP/PS1-EGCG , n = 8), FA ( APP/PS1-FA , n = 8), or EGCG plus FA ( APP/PS1-EGCG/FA , n = 8) for 3 months commencing at 12 months of age (mouse age at sacrifice = 15 months). Data for A and B additionally included WT mice treated in parallel with vehicle ( WT-V , n = 8), EGCG ( WT-EGCG , n = 8), FA ( WT-FA , n = 8), or EGCG plus FA ( WT-EGCG/FA , n = 8). Data for A and B as well as D and E are presented as standard deviations of the means. QPCRs are shown for tumor necrosis factor-α ( TNF -α) or interleukin-1β (IL-1 β) proinflammatory cytokines or for key oxidative stress markers superoxide dismutase 1 ( SOD1 ) or GSH peroxidase 1 ( GPx1 ). Data for A and B are expressed as relative fold over WT-V mice. C , Western blots are shown using anti-Cu/Zn SOD polyclonal antibody ( pAb SOD1 ) or by anti-GPx1 polyclonal antibody ( pAb GPx1 ). Western blots included each mouse ( n = 8 per group), and quantitative data were averaged. Ten μg of protein from each sample was equally loaded per lane. Actin is included as a loading control for each appropriate blot, and densitometry data are shown below each lane. Densitometry data are shown for ratios of SOD1 to actin ( D ) or for GPx1 to actin ( E ). Statistical comparisons for A and B are between-groups but within each mRNA species. Statistical comparisons for D and E are within each protein but between-groups. *, p
Article Snippet: Membranes were then hybridized for 1 h at ambient temperature with primary antibodies as follows: N-terminal anti-APP polyclonal (1:2,000 dilution, IBL); C-terminal anti-presenilin 1 (PS1) monoclonal (1:500 dilution, PS1-loop, Merck Millipore, Darmstadt, Germany); C-terminal anti-sAPP-α monoclonal (1:300 dilution, 2B3; IBL); C-terminal anti-sAPPβ-sw monoclonal (1:100 dilution, 6A1; IBL), N-terminal anti-Aβ(1–16) monoclonal (1:500 dilution, 82E1; IBL); C-terminal anti-BACE1 polyclonal (1:400 dilution, IBL); C-terminal anti-ADAM10 polyclonal (1:1,500 dilution, Cell Signaling Technology, Danvers, MA); anti-Cu/Zn SOD polyclonal (1:3,000 dilution, Enzo Life Sciences, Farmingdale, NY); anti-GPx1 polyclonal (1:4,000 dilution, Boster Biological Technology, Pleasanton, CA); and anti-β-actin monoclonal (1:5,000 dilution, 13E5; Cell Signaling Technology; as a loading control).
Techniques: Mouse Assay, Western Blot
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