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. 2016 Feb 2;14(8):1695–1704. doi: 10.1111/pbi.12530

Figure 4.

Figure 4

Analysis of the effect of a plant signal peptide on necrosis. Improper cleavage of the native signal peptide results in necrosis. (a) Mature TGF‐β1‐specific Western blot analysis with a polyclonal antibody under reducing conditions of plant‐produced SP‐TGF‐β1pc (Tpc) containing the native signal peptide and cSP‐TGF‐β1pc (cTpc) containing the Arabidopsis thaliana chitinase signal peptide at 3 dpi. A molecular weight marker indicates protein size in kDa. As controls, empty vector plant extract (EV) and 5 ng recombinant TGF‐β1 (R5) were used. (b) Leaf necrosis at 5 dpi of SP‐TGF‐β1pc and cSP‐TGF‐β1pc. (c) Yield of SP‐TGF‐β1 and cSP‐TGF‐β1 in crude extracts at 5 dpi as determined by ELISA (n = 3, error bars indicate standard error). Co‐expression of p19 was used in this experiment to further enhance TGF‐β1 expression. (d) Biological activity of 5 ng/mL recombinant TGF‐β1 (rTGF‐β1) and plant‐produced TGF‐β1 (from dpi 5 +  p19) as determined by the induction of luciferase expression in MLEC reporter cells.