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. 2017 Oct 24;19(5):1172–1183. doi: 10.1111/mpp.12595

Figure 2.

Figure 2

Interaction of Agrobacterium tumefaciens C58 VirE2 with AtVIP1 and its homologues. (A) Yeast two‐hybrid interaction assay. LexA‐C58 VirE2 was co‐expressed with Gal4‐AD fused to the indicated tested proteins. The indicated dilutions of cell cultures were plated and grown on non‐selective (+histidine) and selective (–histidine) media in the absence (left) or presence (right) of 0.1 mm 3‐amino‐1,2,4‐triazole (3‐AT). (B) Co‐immunoprecipitation interaction assay. C58 VirE2‐Myc was expressed with GFP‐AtVIP1 and its GFP‐tagged homologues for 3 days in agroinfiltrated Nicotiana benthamiana leaves, immunoprecipitated (IP) with anti‐GFP antibody (top panel), followed by western blot analysis with anti‐GFP or anti‐Myc antibody. To visualize the total amounts of the tested proteins (Input), they were analysed by western blotting with anti‐GFP or anti‐Myc antibody without immunoprecipitation. Lane 1, AtbZIP18; lane 2, AtbZIP52; lane 3, AtbZIP69; lane 4, AtposF21; lane 5, AtbZIP29; lane 6, AtbZIP30; lane 7, NtRSG; lane 8, AtVIP1; lane 9, free green fluorescent protein (GFP). Two independent experiments were performed for each assay with similar results.