Ab13810

Primary antibodies against MARCO (ab239369), TRIF (ab13810), NF‐κB (ab16502), TNF‐α (ab183218), Lamin B (ab133741) and GAPDH (ab181602) as well as the secondary antibody of goat anti‐rabbit were purchased from Abcam. Primary antibody against IκB‐α (4814) was purchased from Cell Signalling Technology. Primary antibody against Toll‐like receptor 4 (TLR4) (AF7017) was purchased from Affinity. PolyG (P4404) and LPS (L2880) was purchased from Sigma.

Cell or tissue proteins were extracted using RIPA solution (P0013C, Beyotime, Shanghai, China) containing protease inhibitors (CW2200S, CWBIO, Beijing, China). After extraction and denaturation of total proteins, 10% gel electrophoresis was used to separate the proteins, and activated PVDF membranes were used for transfer. Subsequently, the blocked-membranes with 5% skim milk powder were incubated with primary antibodies such like anti-TLR4 (1:1000, 14358S, CST, Boston, MA, USA), anti-MyD88 (1:1000, ab219413, Abcam, Cambridge, UK), anti- TIR domain containing adaptor molecule 1 (TRIF) (1:3000, ab13810, Abcam, Cambridge, UK), anti-NF-κB (1:3000, 8242T, CST, Danvers, MA, USA), anti-IκBα (1:3000, 4814T, CST, Danvers, MA, USA), Anti-ATP citrate lyase antibody (ACLY) (1:10000, ab40793, Abcam, Cambridge, UK), phospho- ACLY (1:1000, 4331T, CST, Danvers, MA, USA), and anti- β-actin (1:20000, 81115-1-RR, Proteintech, Chicago, IL, USA) followed by corresponding secondary antibodies (1:6000, 7074/7076, CST, Danvers, MA, USA) at 25 °C for 2 h. The membrane was incubated with ECL reagents (610020-9Q, Qing Xiang, Shanghai, China) and visualized by ImageJ software.

Antibodies specific for TLR2 (ab213676), TLR3 (ab62566), and TRIF (ab13810) were from Abcam. Antibodies specific for TLR4 (HPA049174), TLR5 (HPA015573), TLR7 (HPA059613), and TLR9 (HPA004731) were from Sigma-Aldrich. Antibodies specific for MyD88 (sc-136970) were from Santa Cruz Biotechnology.