Skip to main content
. 2016 Feb 22;6:21651. doi: 10.1038/srep21651

Figure 6. The interaction identification between SGT1 and SRC2-1 or PcINF1 by yeast two- hybrid, co-IP and BiFC.

Figure 6

(a) The interaction confirmation between SGT1 and SRC2-1 by yeast two-hybrid. Yeast transformants expressing the corresponding serial vectors were plated on SD-Leu-Trp-His media containing 50 mM 3-amino-1, 2, 4-triazole (3AT), a competitive inhibitor of the His3p enzyme. The PcINF1/SRC2-1 interaction was used as positive control. (b) The SGT1/SRC2-1 interaction confirmation by co-IP. SRC2-1-HA/SGT1-Flag and GFP-HA/SGT1-Flag protein complexes co-expressed in N. benthamiana leaves (OD595 = 0.8). Anti-HA and anti-Flag antibodies were used to detect SRC2-1 (GFP) and SGT1, respectively. The interaction of GFP-HA and SGT1-Flag were used as a negative control. Coomassie brilliant blue staining of the membrane was to show the equal loading. IB: immunoblotting. (c) BiFC assay of the SGT1/SRC2-1 interaction in N. benthamiana leaves infiltrated with Agrobacterium tumefaciens (OD595 = 0.4). The fluorescent signals were detected using a confocal microscope 48 h after infiltration. Bars = 50 μm.