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. 2017 Feb 14;18(2):409. doi: 10.3390/ijms18020409

Figure 4.

Figure 4

Localization and functional characterization of a nuclear localization signal (NLS) motif for PITG_22798. (A) PITG_22798-GFP fusion protein and control GFP were expressed in N. benthamiana leaves. Photos show confocal images of epidermal cells at 36 h after agroinfiltration. White arrows indicate the nucleus. PITG_22798-GFP is clearly visible in the nucleus (arrow). Left GFP channel, middle bright field channel, right GFP, and bright field channels merged. The white scale bars represent 20 μm. (B) Overview of the mutant construct. nls-PITG_22798-GFP, ΔNLS-PITG_22798-GFP, PITG_22798-GFP and control GFP were transiently expressed in N. benthamiana leaves using agroinfiltration. Confocal microscopy was used to investigate fusion protein distributions. The epidermal cells transformed by nls-PITG_22798-GFP and ΔNLS-PITG_22798-GFP showed the same amount of cytosolic fluorescence as the control GFP. Photos show confocal images of mesophyll cells at 36 h after agroinfiltration. The white scale bars represent 20 µm. The OD600 of the Agrobacterium culture for localization was 0.01. (C) Expression of PITG_22798 and mutants in N. benthamiana leaves by agroinfiltration. The OD600 of the Agrobacterium culture for infiltration was 0.4. Photographs of symptoms were taken 6 d after infiltration. Three repeats show similar results. Each repeat includes 24 infiltration sites on 6 N. benthamiana plants (4 leaves per plant).