Extended Data Fig. 6. Association of Stomagen with ER-family receptors and TMM.

Shown are Co-IP assays of ligand-receptor pairs expressed in N. benthamiana leaves. The ectodomains and membrane-spanning domains of ER, ERL1, and ERL2 fused with GFP were separately expressed in N. benthamiana, and microsomal fractions were incubated with 1 μM Stomagen peptides followed by immunoprecipitation using anti-GFP (αGFP) antibody. Inputs and immunoprecipitates were immunoblotted using anti-GFP (αGFP) or anti-Stomagen (αStomagen) antibodies. Experiments were repeated three times (3 biological replicates). (b) Co-IP of LURE2 peptide fused with hexa-histidine tag (LURE2-His) with N. benthamiana microsomal fractions expressing the ectodomains and membrane-spanning domains of ER and FLS2 fused with GFP, a full-length TMM fused with GFP, or a control, uninoculated leaf sample. Immunopreciptation was performed using αGFP and immunoblotted using αGFP (for detection of receptors) or αHis (for detection of LURE2-His) antibodies. Experiments were repeated twice (two biological replicates). (c) Co-IP of Stomagen peptide with N. benthamiana microsomal fractions expressing the ectodomains and manbrane-spanning domains of ER and FLS2 fused with GFP or a control, uninoculated leaf sample. Immunopreciptation was performed using αGFP and immunoblotted using αGFP (for detection of receptors) or αStomagen antibodies. Experiments were repeated four times (four biological replicates).