FIGURE 3.

Subcellular localization of NbWRKY40 in Nicotiana benthamiana following infection with ToMV. (A) Subcellular localization of NbWRKY40 in response to ToMV infection. Agrobacterium infiltration was used to introduce recombinant plasmid NbWRKY40-GFP + ToMV into healthy N. benthamiana epidermal cells. Healthy leaves infiltrated with an Agrobacterium culture containing NbWRKY40-GFP or infiltrated with an Agrobacterium culture that only contained GFP acted as controls. The confocal microscopy images were captured 2 dpi under bright-field fluorescence to show cell morphology; under dark field to show green fluorescence, indicating localization of the NbWRKY40 protein, and blue fluorescence, indicating nuclei stained blue by 4,6-diamidino-2-phenyl-indole dihydrochloride (DAPI); and under combination fluorescence to show the three images merged. Scale bar, 100 μm. (B) Immunoblot of total protein extracted from the leaves of ToMV-inoculated (+ToMV) and mock-inoculated (–ToMV) plants. Anti-GFP was used to detect GFP fractions. Equal amounts of protein were used for immunoblotting and for staining with Coomassie blue (CBB). (C) Immunoblot of nuclear protein extracted from the leaves of ToMV-inoculated and mock-inoculated plants. Anti-GFP was used to detect GFP fractions. Equal amounts of protein were used for immunoblotting and for staining with CBB.