Anti 4 hne

Thirty teas were purchased from China (for details, see Table 1). 1,1,3,3-Tetranethoxypropane (TMP) (98%) was purchased from Fluka (Seelze, Germany). Bovine serum albumin (BSA) (≥99%) and thioflavin T (ThT) were purchased from Sigma (Saint louis USA). Triglyceride (TG), total cholesterol (TC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) detection kits were purchased from Nanjing Jiancheng Biotechnology Co., Ltd., Nanjing, China. Adenosine triphosphate (ATP) Assay Kit was purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Mouse interleukin-6 (IL-6) and amyloid beta 42 (Aβ42) ELISA kits were purchased from Feiya Biotechnology Co., Ltd. (Haian, China). In addition to anti-GAPDH (Cell Signaling, Boston, MA, USA), the following primary antibodies were used for Western blotting analysis: anti-β-amyloid (Cell Signaling, Boston, MA, USA), anti-multiubiquitin (Medical & Biological Laboratories co. Ltd., Tokyo, Japan), anti-4-HNE (Millipore, Boston, USA), anti-sequestosome-1 (p62) (Epitomics, Burlingame, CA, USA), and anti-RAGE (Santa Cruz Biotechnology, Dallas, TX, USA). Western chemiluminescent horseradish peroxidase substrate was purchased from Millipore. Other reagents used in the experiment are of analytical grade or chromatographic purity grade.

Both EGCG (≥99%) and L-theanine (≥99%) were purchased from Sigma-Aldrich (St Louis, MO, United States). In addition to the secondary antibody (Abcam, CA, United Kingdom), the following primary antibodies were used for western blot analysis: anti-GAPDH, anti-Histone H3, anti-CyclinD1, anti-CyclinB1, anti-IL6, anti-AMPK, anti-mTOR, anti-Sirt1, anti-NF-κB, anti-Nrf2, and anti-KEAP1 (Cell Signaling Technology, Boston, United States), anti-Klotho (Novusbio, Colorad, United States), both RAGE and β-Amyloid (Santa Cruz Biotechnology, San Cruise, United States), anti-p62 (Epitomics, Burlingame, United States), anti-4-HNE (Millipore, Boston, United States), Anti-Multi Ubiquitin mAb (MBL, Tokyo, Japan). Western chemiluminescent horseradish peroxidase substrate (Thermo Fisher Scientific, Waltham, MA, United States), protease inhibitor mixture (Beyotime, Shanghai, China), BCA protein assay reagent (Thermo Fisher Scientific, Waltham, MA, United States), and radioimmunoprecipitation assay (RIPA) lysis buffer (KeyGEN BioTECH, Nanjing, China) were also used in the experiments.