Fig. 6: BIN2 physically interacts with TOE1. | Nature Communications

Fig. 6: BIN2 physically interacts with TOE1.

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

Fig. 6

a A schematic diagram of the TOE1 protein. The TKLVT motif is the classical phosphorylation motif of BIN2. The threonine in red indicates the phosphorylated site. b BIN2 interacts with TOE1 in yeast. BIN2 was fused to pGAD (AD), and TOE1 was fused to pGBK (BD). AD and BD empty vectors were used as the negative controls. c BIN2 interacts with TOE1 in vivo in the nucleus as shown by BiFC assay. BIN2 was fused to cYFP, and TOE1 was fused to nYFP. The recombinant plasmids were transformed into the WT Arabidopsis protoplast. Scale bar = 10 μm. d BIN2 interacts with TOE1 in vitro in a pull-down assay. TOE1-His was incubated with GST or GST-BIN2, and proteins immunoprecipitated with glutathione-agarose beads were detected with an anti-His and an anti-GST antibody, respectively. e BIN2 interacts with TOE1 in vivo in an CoIP assay. Total protein was extracted from Arabidopsis protoplasts containing 35 S::BIN2-MYC, 35 S::3×HA-rTOE1, and the 35 S::BIN2-MYC/35 S::3×HA-rTOE1 combination, and incubated with beads conjugated with an anti-HA. Coimmunoprecipitated proteins were detected with an anti-MYC and an anti-HA antibody, respectively. All experiments were repeated 3 times biologically.

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