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. 2019 Jul 24;10:1684. doi: 10.3389/fmicb.2019.01684

Figure 3.

Figure 3

Transient expression of m460-GFP in the epidermal cells of the N. benthamiana leaves. (A) Schematic of the constructs used for agroinfiltration. The 35S represents CaMV 35S RNA promoter and NOS is the nopaline synthase terminator. NES and NLS indicate PKI nuclear export signal (ELALKLAGL) and SV40 nuclear localization signal (PKKKRKV), respectively. (B) Subcellular localizations of m460-GFP, NES-m460-GFP and NLS-m460-GFP. The constructs were delivered into the five-leaf stage N. benthamiana plants continuously grown at 25°C via agroinfiltration. Moreover, the plasmid bearing m460-GFP was introduced into plants pre-grown at 32 or 18°C for 3 days before agroinfiltration. The infiltrated plants were further grown at 25, 32, or 18°C for 60 h, after which the fluorescence was viewed by confocal microscopy. H2B-RFP was used as a marker for the nucleus. The sublocalization of GFP alone at 25°C was comparable to that at 32 or 18°C (data not shown). All experiments were repeated twice with similar results.