Rabbit anti trem 1

Immunoblot analysis was performed as described earlier [13 (link)], using the following antibodies: rabbit anti-TREM-1 (Abcam, USA), rabbit anti-MyD88 (Cell Signaling, USA), rabbit anti-TRIF (Thermo Fisher Scientific, USA), mouse anti-Syk (Abcam, USA), rabbit anti-p-Syk (Thermo Fisher Scientific, USA), rabbit anti-DAP12 (Cell Signaling, USA), rabbit anti-AKT (Cell Signaling, USA), rabbit anti-p-AKT (Cell Signaling, USA), mouse anti-TLR4 (Santa Cruz, USA), rabbit anti-TLR2 (EMD Millipore, USA), rabbit anti-p38 (Cell Signaling, USA), rabbit anti-p-p38 (Cell Signaling, USA), rabbit anti-ERK (Cell Signaling, USA), rabbit anti-p-ERK (Cell Signaling, USA), rabbit anti-IκB (Cell Signaling, USA), rabbit-anti-p-IκB (Cell Signaling, USA), mouse anti-C/EBP (Santa Cruz, USA), mouse anti-β-actin (Santa Cruz, USA), and rabbit anti-4G10 (Sigma-Aldrich, USA). Either horseradish peroxidase-conjugated goat anti-rabbit antibody or goat anti-mouse antibody (Cell Signaling, USA) was used as a secondary antibody. R406 was purchased from Selleckchem.

Western blot was performed as previously described.¹⁶ The right brain hemispheres were homogenized in RIPA (Santa Cruz Biotechnology) and centrifuged at 14,000 g at 4°C for 30 min. Equal samples were loaded on an SDS‐PAGE gel, and then the proteins were electrophoresed and transferred to a nitrocellulose membrane. The membranes were incubated overnight at 4°C with the primary antibodies as followed: rabbit anti‐TREM‐1 (1:1000, Abcam), rabbit anti‐SYK (1:1000, Abcam), rabbit anti‐β‐Catenin (1:3000, Abcam), rabbit anti‐phospho‐β‐Catenin (1:1000, Cell Signaling Technology), rat anti‐ ZO‐1 (1:500, Santa Cruz Biotechnology), rabbit anti‐Claudin‐5 (1:1000, Abcam), and mouse anti‐β‐actin (1:3000, Santa Cruz Biotechnology). The respective secondary antibody (1:3000, Santa Cruz Biotechnology) were incubated at room temperature for 2 h. Immunoblots were probed with an ECL Plus chemiluminescence reagent kit (Amersham Biosciences) and visualized with an imaging system (Bio‐Rad, VersaDoc, model 4000). Relative density was analyzed using Image J software.