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. 2014 Jun 27;14:59. doi: 10.1186/1472-6750-14-59

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Copyright © 2014 Pereira et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Accumulation and activity of AnPGI 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.