Fig. 5

SlNAP1 interacts with SlGID1 in vitro and in vivo. A BiFC assays. Full-length SlNAP1 and SlGID1 were fused to the N-terminal part and the C-terminal part of YFP, respectively. Constructs were transformed to Agrobacterium tumefaciens strain GV3101, and were then injected into 4-week-old tobacco leaves. The YFP fluorescence was observed under a confocal laser scanning microscope after incubating at 22 °C for 24–48 h. Bars = 50 μm. B Y2H assays. The full-length SlGID4 was fused with the activation domain (pGADT7-SlGID4) and the full-length SlNAP1 was fused with the binding domain (pGBK7-SlNAP1). Transformed yeast cells were grown on SD-Leu-Trp, or SD- Leu-Trp-His-Ade media. These experiments were performed three times with similar results, and a representative picture was shown. C and D LUC assays. CDS of SlNAP1 (with no stop codon) was cloned into pCAMBIA1300-nLUC, and the CDS of SlGID1 was cloned into the pCAMBIA1300-cLUC vector. The constructs were transformed into Agrobacterium tumefaciens strain GV3101, and A. tumefaciens was mixed (1:1, v/v) and coinfiltrated into tobacco, and luminescence was observed in optical in vivo imaging and was analyzed by PlantView. BiFC, bimolecular fluorescence complementation; DAPI 4, 6-diamidino-2-phenylindole, Y2H yeast two-hybrid, LUC Luciferase