Fig. 7: Yeast two-hybrid assay for the physical interaction between CmSI and CmPIN2. | Horticulture Research

Fig. 7: Yeast two-hybrid assay for the physical interaction between CmSI and CmPIN2.

From: Melon short internode (CmSi) encodes an ERECTA-like receptor kinase regulating stem elongation through auxin signaling

Fig. 7

a The protein interaction was examined using various combinations of prey and bait vectors. The vector pPR3-N and pTSU2-App was used as a negative control, and the interaction between pTSU2-APP and pNubG-Fe65 was used as a positive control. Yeast cells were grown on SD/-Leu-Trp medium and interactions were confirmed by an SD/-Leu-Trp-His-Ade-X-a-GAL assay on medium. Dilutions (1 and 10−1) of saturated cultures were spotted onto the plates. Yeast two-hybrid assay showing that CmSI rather than Cmsi interacted with CmPIN2 by growth on SD/-Leu/-Trp/-His/-Ade/X-α-gal plates. b BiFC analysis of the physical interaction between CmSI and Cmsi (fused with the C-terminal fragment of YFP) and CmPIN2 (fused with the N-terminal fragment of YFP). INDEHISCENT (IND)-YFPC and SPATULA (SPT)-YFPN were used as positive controls. CmPIN2-YFPN and the empty YFPC were used as negative controls. Different combinations of the fused constructs were co-expressed in leaves of Nicotiana tabacum, and the cells were then visualized using confocal microscopy

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