Figure 4

Accumulation and activity of An PGI and its fusions with ELP and HFBI targeted to the ER and vacuole. A) Western blot analysis of AnPGI and its fusions targeted to the ER or the vacuole. Each lane was loaded with 5 μg TSP from three replicates, and detected with anti c-myc antibody. B) The effect of fusion partner on the accumulation of AnPGI. C) A N. benthamiana leaf disc agroinfiltrated with AnPGI and its fusions was incubated in 50 mM of sodium acetate at 50°C for 24 h and used to analyze self hydrolysis and release of reducing sugars using the dinitro salicylic acid (DNS) method. D) The specific activity against polygalacturonic acid was used to analyse the effect of ELP and HFBI fusions on AnPGI activity. Accumulation results represent the average of AnPGI levels in five different plants, and the release of reducing sugars and protein activity were determined in triplicate. Reducing sugar concentration was normalized using the assay results for wild-type plants. Error bars represent ± standard error.