Extended Data Fig. 6: Phosphorylated MtLICK1/2 are less efficient in forming a complex with MtLYK3.

a,b, Western blot detection of MtLICK1^WT and mutants in yeast. Western blot membrane was immunoblotted with a-HA and a-Myc to indicate the amount of MtLICK1 or MtLYK3_CD respectively. c,d, Western blot detection of MtLICK2^WT and mutants in yeast. Western blot membrane was immunoblotted with a-HA and a-Myc to indicate the amount of MtLICK2 or MtLYK3_CD respectively. e,f, MtLICK1/2^{WT or 3D} -Flag or Flag was expressed in Medicago stable transgenic plants (pMtLYK3:gMtLYK3-GFP) hairy roots transformed with pLjUBQ:MtLICK1/2^{WT or 3D} or pLjUBQ:Flag by hairy root transformation. Co-IP assay was performed with a-GFP. Flag was expressed in Medicago stable transgenic plants (pMtLYK3:gMtLYK3-GFP) by hairy root transformation serves as control. The histograms right show a statistical analysis of the relative signal intensity based on three repeated Co-IP assay data. Data are mean ± SEM. Statistical analysis was performed using Two-sided Student’s t-test compared to MtLICK1^WT or MtLICK2^WT. g,h, MtLYK3-HA and GFP-MtLICK1/2^WT or GFP-MtLICK1/2^3D-Flag were transiently co-expressed in N. benthamiana leaves transformed with pLjUBQ:MtLYK3-HA, p35S:GFP-MtLICK1/2^WT and p35S:GFP-MtLICK1/2^3D-Flag. Co-IP assay was performed with a-GFP. GFP and HA or GFP and MtLYK3-HA were transiently co-expressed in N. benthamiana leaves serve as control. The histograms right show a statistical analysis of the relative signal intensity based on three repeated Co-IP assay data. Data are mean ± SEM. Statistical analysis was performed using Two-sided Student’s t-test compared to MtLICK1^WT or MtLICK2^WT. All the experiments were repeated three times with similar results.

Source Data