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. 2020 Sep 11;9(9):1186. doi: 10.3390/plants9091186

Figure 3.

Figure 3

Protein–protein interaction studies involving LTSF1 and LTSF2. (A) Yeast two-hybrid assays analyzing the interactions between LTSF1/2 and SKP1. The LTSF1, LTSF2, and SKP1 coding sequences were cloned into the pGAD-T7 and pGBK-T7 vectors. Yeast AH109 cells transformed with the designated plasmid combinations were grown on SD/-Leu/-Trp and SD/-Leu/-Trp/-His/+3-aminotriazol (3-AT) media. (B) β-galactosidase (GUS) activity assay. Error bars indicate SD of five biological replicates of relative GUS activity assay. The asterisk indicates a significant difference compared with the negative construct (p < 0.05). (C) Bimolecular fluorescence complementation assay of the interactions between pepper LTSF1/2 and SKP1. The LTSF1, LTSF2, and SKP1 coding sequences were cloned into the pSPYCE and pSPYNE vectors. The designated plasmid combinations were co-bombarded into onion epidermal cells for the transient expression of yellow fluorescent protein (YFP), and the fluorescence signals were visualized using a laser-scanning confocal microscope. PC and NC indicate positive and negative controls.