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. 2018 May 15;69(16):4003–4015. doi: 10.1093/jxb/ery182

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© The Author(s) 2018. Published by Oxford University Press on behalf of the Society for Experimental Biology.

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 reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 2. — In planta interaction of CIPK9 and AP2C1 by fluorescence resonance energy transfer (FRET) analysis. (A) Fusion constructs of AP2C1, KIM, and K2 fragments with cyan fluorescent protein (CFP) and of CIPK9 with yellow fluorescent protein (YFP) were co-inoculated in Nicotiana benthemiana cells. The cells that showed expression of both CFP and YFP were targeted for FRET analysis according to the acceptor-bleaching protocol. Representative interactions are shown for different FRET combinations. Arrows indicate the region of interest selected for detailed analysis and calculation of FRET efficiency in individual cells. Scale bars are 20 µm. (B) FRET efficiency, calculated as the mean (±SD) of seven different cells. For the negative control, the efficiency was calculated for cells transformed with vectors containing CFP and YFP only (CFP X YFP). *P<0.05 compared with the negative control.