Figure 5. Characterisation of TLR-stimulants present in human soluble faecal extract.

(A) Human SFE (10 µg/ml) was treated with immobilised lipase for 18 h at 37°C, and then assayed for potential to stimulate TLR2 signalling in HEK-293-TLR2 cells. (B) Human SFE (10 µg/ml), or 100 ng/ml E. coli LPS, was treated with 10 µg/ml polymyxin-B or 1 µg/ml lipid-IVa for 10 minutes before addition to HEK-293-TLR4 cells to measure capacity to induce TLR4-signalling. (C) Human SFE (10 µg/ml), or 500 ng/ml S. typhimurium flagellin was treated with proteinase-K for 4 h before enzyme denaturation at 80°C for 10 minutes, before application to HEK-293-TLR5 cells to measure capacity to induce TLR5-signalling.