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

From: Competitive binding of antagonistic peptides fine-tunes stomatal patterning

Extended Data Figure 6

a, Shown are co-immunoprecipitation 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 (anti-GFP) antibody. Inputs and immunoprecipitates were immunoblotted using anti-GFP (anti-GFP) or anti-Stomagen (anti-Stomagen) antibodies. Experiments were repeated three times (three biological replicates). b, Co-immunoprecipitation 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. Immunoprecipitation was performed using anti-GFP and immunoblotted using anti-GFP (for detection of receptors) or anti-His (for detection of LURE2–His) antibodies. Experiments were repeated twice (two biological replicates). c, Co-immunoprecipitation of Stomagen peptide with N. benthamiana microsomal fractions expressing the ectodomains and membrane-spanning domains of ER and FLS2 fused with GFP or a control, uninoculated leaf sample. Immunoprecipitation was performed using anti-GFP and immunoblotted using anti-GFP (for detection of receptors) or anti-Stomagen antibodies. Experiments were repeated four times (four biological replicates).