Fig. 1: Equilibrium binding with a radiolabelled LCO shows high affinity LCO and CO binding to PhLYK9 and BdLYR2.

a Images of epidermal cells from N. benthamiana leaves expressing the indicated proteins tagged with YFP. Scale bars represent 20 µm. b Specific binding of LCO-V(C18:1,NMe,³⁵S) to PhLYK9 and BdLYR2. Bars represent the mean specific LCO-binding/µg membrane proteins. n = two replicates on two independent batches of membrane fractions containing the indicated proteins or from untransformed leaves. Immunodetection with anti YFP antibodies in 10 µg of the indicated membrane fractions. c Affinity of PhLYK9 and BdLYR2 for LCO-V(C18:1,NMe,S). Scatchard plots of cold saturation experiments using membrane fractions containing the indicated proteins and a range of LCO-V(C18:1,NMe,S) concentrations as competitor. The plots are representative of experiments performed with two independent batches of membrane fractions. d Specificity of PhLYK9 and BdLYR2 LCO-binding sites for LCO versus other GlcNAc-containing molecules. Bars represent the mean percentage of specific LCO-V(C18:1,NMe,³⁵S) binding competition in the presence of 2 μM of the indicated competitors, except for PGN fragments which were used at 10 mg/L. n = technical replicates. LCO is LCO-V(C18:1,NMe,S). e Affinity of PhLYK9 and BdLYR2 for CO4 and CO8. Competitive inhibition of the radiolabelled LCO-V(C18:1,NMe,S) binding to membrane fraction containing the indicated proteins, using a range of concentrations of unlabeled CO4 or CO8 as competitors. f Affinities of PhLYK9 and BdLYR2 LCO-binding sites for the indicated ligands, deduced from saturation and competitive inhibition experiments. Statistical differences (p < 0.05) were calculated using a pairwise two-sided Student’s t-test in b, d.