Fig. 1. TLR dependence of cellular responses to the LPS from R. species. (A) HEK293 cells transfected with the TLR2 and/or TLR6 expression vectors together with a pELAM luciferase reporter plasmid and pRL-TK were exposed 24 hours later to the Rhizobium species Sin 1 LPS (100 ng/ml) for 8 hours. TLR-dependent activation was measured by the dual-luciferase reporter assay. (B) BMGs from C3H/HeOU and TLR2^-/- mice were incubated with lipid A fractions (1 µg/ml) isolated from the LPSs of Bordetella pertussis and Rhizobium species Sin-1. Cell lysates (10⁵ cells in 1% CHAPS, supplemented with a mixture of protease inhibitors) were analyzed for CD14 by SDS-PAGE and western blotting.