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. 2016 Jan 22;7:4. doi: 10.3389/fpls.2016.00004

FIGURE 3.

FIGURE 3

Subcellular localization and transactivation activity of the ONAC022 protein. (A) ONAC022 is localized in nucleus. Agrobacteria carrying pFGC-Egfp-ONAC022 or pFGC-Egfp empty vector were infiltrated into leaves of Nicotiana benthamiana plants expressing a red nucleus marker protein RFP–H2B and leaf samples were collected at 24 h after infiltration for observation under a confocal laser scanning microscope. Images were taken in dark field for green fluorescence (left) and red fluorescence (middle right), white field for cell morphology (middle left) and in combination (right), respectively. (B) DNA binding activity of ONAC022. Wild type version of the cis-element sequence (wNACRS) and a mutated version (mNACRS) were used. Electrophoretic mobility shift assays were performed using the recombinant GST-fused ONAC022 protein. Biotin-labeled wNACRS and mNACRS probe and biotin-labeled wNACRS probe in combination with unlabeled wNACRS or mNACRS probe were incubated with GST-fused ONAC022 protein or a purified GST preparation as a negative control. Specific DNA-protein complexes and free probes are indicated by the arrowheads on left. (C) ONAC022 has transactivation activity. Yeast cells carrying pBD-ONAC022, pBD-ONAC022ΔC, pBD-ONAC022ΔN or pBD empty vector (as a negative control) were streaked on SD/Trp- plates (left) or SD/Trp-His- plates supplemented with x-α-gal (right) for 3 days at 28°C.