Fig. 4: BIN2 physically interacts with SPL9. | Nature Communications

Fig. 4: BIN2 physically interacts with SPL9.

From: Coordinated regulation of vegetative phase change by brassinosteroids and the age pathway in Arabidopsis

Fig. 4

a BIN2 interacts with SPL9 in yeast. BIN2 was fused to pGAD (AD), and SPL9 was fused to pGBK (BD). AD and BD empty vectors were used as the negative controls. DDO (SD/-Leu/-Trp), QDO (SD/-Leu/-Trp/-Ade/-His). b BIN2 interacts with SPL9 in vivo in the nucleus as shown by bimolecular fluorescence complementation (BiFC) assay. BIN2 was fused to cYFP, and SPL9 was fused to nYFP. The recombinant plasmids were transformed into the WT Arabidopsis protoplast. The nucleus was stained with hoechest 33342. Scale bar = 10 μm. c BIN2 interacts with SPL9 in vitro in a pull-down assay. SPL9-His was incubated with GST or GST-BIN2 protein, and the proteins immunoprecipitated with glutathione-agarose beads were detected with an anti-His and an anti-GST antibody, respectively. d BIN2 interacts with SPL9 in vivo in an CoIP assay. Total protein was extracted from Nicotiana benthamiana leaves infiltrated with Agrobacterium containing 35 S::3×FLAG/35 S::3×FLAG-rSPL9, and 35 S::BIN2-GFP/35 S::3×FLAG-rSPL9 combinations, and incubated with anti-FLAG beads. Coimmunoprecipitated proteins were detected with an anti-FLAG and an anti-GFP antibody, respectively. All experiments were repeated 3 times biologically.

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