Fig. 4

Eae1 localization in plant cells. a Microscopic analysis of GFP, Eae1-GFP, and Eae1_NbPR1-SP-GFP in N. benthamiana. White arrows indicate the chloroplast-localized Eae1-GFP. Scale bars: 50 µm. b Microscopic analysis of the Eae1-GFP and Eae1_NbPR1-SP-GFP in the protoplasts isolated from the N. benthamiana leaves. Scale bars: 50 µm. c The His-tagged Eae1 (Eae1-His) was incubated with the protoplasts of H. brasiliensis mesophyll cells for 6 h, and then the chloroplast faction was extracted for western blotting. Input, the His-tagged proteins detected pre-incubation; Protoplast, the total proteins of the protoplasts after incubation; Chloroplast, chloroplast fractions extracted from the protoplasts after incubation. Anti-RuBisCO was used as a loading control. A truncated GFP protein (168 aa) fused with His-tag (GFP₁₆₈-His) and Eae1_NbPR1-SP-His were used as controls. The experiments were conducted twice with similar results