Fig. 5
The interaction between Eae1 and HbAIG1. a Yeast two-hybrid (Y2H) assays detecting the interaction between the Eae1 and HbAIG1. Positive colonies displayed a blue-color on the SD-LWHA medium with X-α-GAL. The combination of AD-T and BD-p53 was used as a positive control. -LW, SD medium lacking Leu and Trp; -LWHA, SD medium lacking Leu, Trp, His, and Ala. Red asterisk indicates the combination of Eae1 and HbAIG1. b Bimolecular fluorescence complementation (BiFC) verifies the interaction between Eae1 and HbAIG1 in N. benthamiana. Scale bars: 100 µm. White arrows indicate the represented interacted region. c The interaction between Eae1-GFP and HbAIG1-FLAG was determined by co-immunoprecipitation (Co-IP) assays. Input, total proteins extracted from N. benthamiana leaves co-expressing two tested proteins. IP, the proteins eluted from anti-GFP beads. Red asterisks indicate the interested bands. d The interaction between Eae1-His and HbAIG1-GST was determined by the pull-down assay. A truncated GFP protein (168 aa) fused with His-tag (GFP168-His) was used as controls. Red asterisks indicate the interested bands. The experiments were conducted twice with similar results