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. 2022 Mar 21;13:1524. doi: 10.1038/s41467-022-29196-6

Fig. 5. In planta association of the engineered full-length RGA5 receptors with MAX effectors.

Fig. 5

The full-length RGA5 protein and related m1, m2, and m1m2 variants fused to YFP were transiently co-expressed in N. benthamiana leaves with the indicated HA-tagged effectors (without signal peptides). The HMA domain of Pikp-1 (fused to YFP) was used as a control as well as YFP alone and the M. oryzae effector PWL2. Proteins were extracted 48 h after infiltration, separated by gel electrophoresis, and tagged proteins were detected in the extract (input) and after immunoprecipitation with anti-GFP beads (IP-GFP) by immunoblotting with anti-HA (α-HA) and anti-GFP (α-GFP) antibodies. In the IP-GFP/α-GFP panel, yellow and white asterisks indicate the YFP:Pikp-1_HMA and YFP proteins respectively, while blue asterisks show the YFP:RGA5 wild type and mutant proteins. Ponceau staining shows equal protein loading in the input. The experiment was carried out twice with identical results.