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. 2014 Aug 30;14:231. doi: 10.1186/s12870-014-0231-5

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© Garcia-Molina et al.; licensee BioMed Central Ltd. 2014

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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Biochemical analysis of SPL7 subcellular localization and processing. (a) Total protein extracts from tobacco leaves transiently expressing different GFP-tagged SPL7 versions were subjected to biochemical fractionation, as described in Experimental Procedures and analysed by Western blotting with antibodies against GFP (α-GFP). Antibodies against the organelle markers TPR7 (α-TPR7), PEPC (α-PEPC) and histone H3 (α-H3) were used to validate the fractionation. M, microsomes; C, cytosol; N, nucleus. Main bands are highlighted with an asterisk. (b) Proteolytic processing of transiently expressed GFP::SPL7 and SPL7::GFP by Western blotting. WT, protein extract of non-transformed tobacco leaves. Sizes of molecular-weight markers run in the same gels are shown alongside the blots in (a-b) according to manufacturer’s indications for 10% SDS-PAGE gels.