Fig. 7
Eae1 can reduce HbSAMS5 protein abundance. a Western blotting shows that co-expression of Eae1-GFP and HbSAMS5-FLAG resulted in a relatively low HbSAMS5-FLAG content in N. benthamiana. RuBisCO stained with CBB was used as a loading control. The red arrow indicates the lane with the loading of plant proteins containing Eae1-GFP and HbSAMS5-FLAG. GUS370-FLAG, a truncated β-glucuronidase (GUS) protein (370 aa) fused with FLAG. b In vitro protein incubation assay, Eae1-His protein promoted the destabilization of HbSAMS5-GST when leaf cytosolic extract was added. The proteins were analyzed by western blotting pre- and post-incubation. RuBisCO stained with CBB represents the loading of leaf cytosolic extract. The red arrow indicates the lane with the loading of plant proteins containing Eae1-His and HbSAMS5-GST. c Eae1-His did not induce the destabilization of HbSAMS5-GST without the addition of leaf cytosolic extract following incubation. RuBisCO stained with CBB represents the loading of leaf cytosolic extract. d HbSAMS5-GST exhibited a decrease in response to increasing concentrations of Eae1-His. RuBisCO stained with CBB represents the loading of leaf cytosolic extract. Similar results were obtained from two independent experiments, and red asterisks indicate the interested bands. e Eae1NbPR1-SP-GFP did not suppress INF1-induced HR. The values represent the leaves with HR in 9 tested leaves from 3 independent experiments