Anti psd 95

Lipopolysaccharide and caffeine were purchased from Sigma-Aldrich Chemicals Company (St. Louis, MO, USA). The antibodies used in the Western blot and immunofluorescence studies were anti-Nrf2 (sc-722), anti-HO1 (sc-136,961), anti-synaptosomal-associated protein 23 (SNAP-23) sc-374,215), anti-PSD-95(sc-71,933), anti-TNF-α (sc-52,746), TLR4 (sc-16240), anti-Bax(sc-7480), anti-Bcl2 (sc-7382), anti-p-NF-kB (sc-36,548), anti-Iba-1 (sc-32,725), anti-Glial fibrillary acidic protein (GFAP; sc-33,673), and anti-β-actin (sc-7,778) (Santa Cruz Biotechnology, Dallas, TX, USA). In addition, the anti-Cleaved Caspase-3 #9664) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Other primary antibodies were diluted in 1× TBST (1:1000), and secondary anti-mouse horseradish peroxidase (HRP)-conjugated (Promega Ref#W402) and anti-rabbit HRP-conjugated (Promega Ref# W401) antibodies were diluted 1:10,000 in 1× TBST(Sigma, Fitchburg, WI, USA). For the confocal microscopic studies, secondary fluorescent antibodies goat anti-mouse (Ref# A11029) and goat anti-rabbit (Ref# 32732) diluted in 1× PBS were used.

Fisetin (Lot#SLBF3913V) and d-galactose (sc-202564) were procured from Sigma-Aldrich Chemical Co. (St. Louis, MO, United States). The drugs were dissolved in 0.1% dimethyl sulfoxide (DMSO) and the final volume was adjusted with normal saline (0.9% saline). The control mice were injected with normal saline.
The antibodies used in the current studies are: SIRT1 (sc-74465), anti-Nrf2 (sc-722), anti-HO-1 (sc-136,961), P-JNK (sc-625), anti-p-NF-κβ (sc-136,548), anti-Iba-1 (sc-32,725),anti-GFAP (sc-33673) anti-interleukin (IL-1β) (sc-32,294), anti-tumor necrosis factor-α (TNF-α) (sc-52,746), NOS-2 (sc-651), anti-Bax (sc-7480) anti-Bcl2 (sc-7382), cleaved Caspase-3 (sc-7272), anti-PARP-1 (sc-8007), anti-PSD-95 (sc-71,933), anti-synaptosomal-associated protein 23 (SNAP-25), and anti-β-actin (sc-47,778) (Santa Cruz Biotechnology, Dallas, TX, United States). The primary antibodies were diluted in 1× TBST (1:1,000), and secondary anti-mouse HRP (horseradish peroxidase) conjugated (Promega Ref# W402) and anti-rabbit HRP conjugated (Promega Ref# W401) were diluted 1:10,000 in 1 M TBST (Promega, Fitchburg, WI, United States); the secondary antibodies (anti-mouse Ref# A11029 and anti-rabbit Ref# 32,732) used in the immunofluorescence studies were diluted in 1:100 in 1 M PBS.

The antibodies used in the Western blot and immunofluorescence studies were anti-Nrf2 (sc-722), anti-HO1 (sc-136,961), anti-synaptosomal-associated protein 23 (SNAP-23) (sc-374,215), anti-PSD-95 (sc-71,933), anti-Syntaxin (sc-12,736), anti-tumor necrosis factor-α (TNF-α) (sc-52,746), anti-PARP-1 (sc-8007), TLR4 (sc-16240), Synaptophysin (sc-17750), anti-interleukin (IL)-1β (sc-32,294), anti-Bax (sc-7480), anti-Bcl2 (sc-7382), anti-p-NF-κB (sc-136,548), anti-Iba-1 (sc-32,725), anti-Glial fibrillary acidic protein (GFAP; sc-33,673), and anti-β- actin (sc-47,778) (Santa Cruz Biotechnology, Dallas, TX, USA). In addition, the anti-Cleaved Caspase-3 (#9664) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). For Syntaxin and β-actin, the antibodies were diluted in TBST (1:10000) (Santa Cruz Biotechnology, Dallas, TX, USA). Other primary antibodies were diluted in 1x TBST (1:1000), and Secondary anti-mouse HRP (Horseradish peroxidase) conjugated (Promega Ref# W402) and anti-rabbit HRP conjugated (Promega Ref# W401) were diluted 1:10,000 in 1× TBST and were purchased from Promega, (Fitchburg, WI, USA). TAK242, Resatorvid (CAS 243984-11-4), the specific inhibitor of TLR4. For the confocal microscopic studies, the secondary fluorescent antibodies used were goat anti-mouse (Ref# A11029) and goat anti-rabbit (Ref# 32732) diluted in 1× PBS.

Anti-GluK3 antibody raised in rabbit against C-terminal 17 amino acid residues of mouse GluK3 (903–919, NM_001081097) was used for immunohistochemistry, immunoprecipitation, and western blotting. The rabbit polyclonal antibodies anti-GluK2 (Synaptic Systems, 180 003), anti-GluK4^{41 (link)}, and anti-GluK5 (Millipore, 06-315) were used for immunoprecipitation and western blotting. The specificities of these antibodies have been determined previously^{29 (link)}. Anti-GluN1 (BD Biosciences, 556308), anti-GluN2A (BD Biosciences, 612286), anti-GluN2B (BD Biosciences, 610416), anti-GluA1 (Frontier Institute, Japan, MSFR102270), anti-GluA2 (Millipore, MAB397), anti-GluA3^{44 (link)}, anti-PSD-95 (Santa Cruz Biotechnology, SC32290), anti-D1R (Frontier Institute, Japan, MSFR101030), and anti-D2R (Frontier Institute, Japan, MSFR101060) were used for western blotting.