Figure 3.

Subcellular localization and necrotizing activity of SsNE2 with its native SP and tagged with various targeting signals upon Agrobacterium tumefaciens-mediated expression in Nicotiana benthamiana leaves. (A) Schematic representation of the constructs showing native signal peptide (SP), signal peptide derived from A. thaliana Pathogenesis-related Protein 1 (SP^PR1), SsNE2 protein (SsNE2), nuclear localization signal (NLS), nuclear export signal (NES), calcineurin B–like protein 1 myristoylation signal (CBL1), endoplasmic reticulum (ER) retention signal (KDEL) and green fluorescent protein (GFP). (B) Analysis of necrosis produced by each SnNE2 protein tagged with various targeting signals in N. benthamiana leaves. (C) Microscopic localization of each SsNE2 protein fused to GFP and tagged with various targeting signals. To confirm ER localization, constructs expressing effector proteins were co-infiltrated with a construct expressing an mCherry protein tagged with an ER retention signal (mCherry::KDEL).