Fig. 5.

Minimal autoactive domains of MLA10, Sr33, and Sr50 self-associate in planta. (A) MLA10, Sr33, and Sr50 protein fragments fused to HA or CFP were transiently expressed in N. benthamiana. The autoactive MLA10^1-160:CFP, Sr33^1-160:CFP, and Sr50^1-163:CFP constructs were used as positive controls. Cell death was visualized 5 d after infiltration. Equivalent results were obtained in three independent experiments. (B) Indicated proteins, transiently expressed in N. benthamiana leaves, were extracted 20 h after infiltration and analyzed by immunoblotting with anti-HA (α-HA) and anti-GFP antibodies (α-GFP) (Input). Proteins were immunoprecipitated with anti-GFP beads (IP-GFP) and analyzed by immunoblotting with anti-GFP and anti-HA antibodies. RGA4 (CC domain):CFP fusion was used as a control for specificity. Sr50^1-163 was used as a positive control (25). Ponceau staining of the RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase) large subunit shows equal protein loading.