Ab126758

Retinoic acid was obtained from Sigma-Aldrich China (Shanghai, China; R2625). Rapamycin was purchased from MedChemExpress (Shanghai, China; HY-10219). CTB-Alexa 488 and CTB-Alexa 555 were bought from BrainVTA (Wuhan, Hubei, China). The following antibodies were used in western blot: anti-CD82 (ab66400 & ab135779; Abcam; 1:1000); anti-synaptophysin (ab14692; Abcam; 1:1000); anti-p70S6k (2708T; Cell Signaling Technology; 1:1000); anti-p-p70S6k(9234T;Cell Signaling Technology; 1:1000); anti-β-actin (sc-47778; Santa Cruz; 1:1000); anti-TRAF2 (ab126758; Abcam; 1:1000); anti-Raptor (20984-1-AP; Proteintech; 1:1000).The following antibodies were used in immunofluorescence: anti-CD82 (ab66400; Abcam; 1:200); anti-Tuj1 (801201; Biolegend; 1:1000); anti-Iba1(ab48004;Abcam;1:100); anti-GAFP(ab4674; Abcam;1:100); anti-mCherry (ab167453; Abcam; 1:100); anti-synaptophysin (ab14692; Abcam; 1:200); anti-pS6(Ser 240/244)(5364T; Cell Signaling Technology; 1:1000); anti-TRAF2 (ab126758; Abcam; 1:400); anti-Raptor (20984-1-AP; Proteintech; 1:1000); anti-Lamp1 (15665, Cell Signaling Technology; 1:500).
Anti-β-Amyloid (2450; Cell Signaling Technology; 1:100) was used in immunohistochemistry.

The protein was extracted with RIPA Lysis buffer (Boster, AR0102-100, Wuhan, China) and quantified by ultrastructure nucleic acid protein tester (Analytik Jena, Jena, Germany). Equal amounts of protein were separated by SDS-PAGE, immunoblotted with antibodies IRE1α (Proteintech, 27528-1-AP, Wuhan, China), anti-TRAF2 antibody (Abcam, ab126758, Cambridge, UK), anti-IKK-β antibody (Abcam, ab124957, Cambridge, UK), IκB-α antibody (CST, #4812, Boston, MA, USA), NF-κBp65 antibody (CST, #8242, Boston, US), ASK1 antibody (Proteintech, 67072-1-Ig, Wuhan, China), anti-JNK1 antibody (Abcam, ab199380, Cambridge, UK), and GAPDH antibody (Proteintech, 60004-1-IG, Wuhan, China). Antibody binding was detected using horseradish peroxidase-conjugated anti-rabbit IgG/anti-mouse IgG antibody (Proteintech, SA00001-1/SA00001-2, Wuhan, China) and visualized with Chemiluminescence Solution (Abbkine, BMU102-CN, Wuhan, China).

Raw264.7 cells and M2 macrophages were lysed using the RIPA lysis buffer on ice. Proteins were extracted from M2-exos using the exosome protein extraction kit (EZBioscience, Roseville). Afterwards, an equal amount of proteins were run to separate using the 10% SDS-PAGE and transferred to PVDF membranes (Millipore, Billerica). The membranes were blocked using 5% skim milk and incubated the following primary antibodies at 4 °C overnight. The antibodies (Abcam, Cambridge) including anti-TSG101 (ab125011, 1/1000), anti-CD9 (ab263019, 1/1000), anti-TNF-α (ab183218, 1/1000), anti-TRAF2 (ab126758, 1/1000), anti-FADD (ab124812, 1/1000), and anti-GAPDH (ab9485, 1/2500). A secondary antibody (ab97051, 1/10,000) was continued to incubate with the membranes at room temperature for 1 h. The bands were visualized using an ECL western blotting substrate kit (Abcam).