Immunoblot Analysis of Inflammatory Signaling

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.