Expression of endogenous ligands for TLR2. (A) Expression of biglycan in AD-TLR2KO mice increased significantly compared with that in WT, AD, and TLR2KO mice (p<0.05). (B) HMGB1 in the four groups did not show a significant difference (p>0.05).

Western Blot: HMGB1/HMG-1 Antibody [TA301448] - Total protein from SHSY-5Y, MCF7, Neuro2A and HeLa was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-HMGB1 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Flow Cytometry: HMGB1/HMG-1 Antibody [TA301448] - An intracellular stain was performed on RH-30 cells with TA301448F (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to FITC.