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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. 4. — Dephosphorylation of auto-phosphorylated CIPK9 by AP2C1. (A) Autoradiogram showing detection of signals after adding different combination of GST-CIPK9 and GST- AP2C1 proteins. Lanes of interest were sliced from same autoradiogram/gel and placed together for easy representation of the data. Each sliced part is clearly demarcated by border lines. (B) To determine the dose-dependent activity of AP2C1, the concentration of GST-AP2C1 was increased gradually from 0–250 ng while GST-CIPK9 was kept constant at 500 ng in all the reactions. The autoradiogram shows the gradual decrease in the auto-phosphorylation signal of CIPK9. (C) A gradual increase in the concentration of GST-AP2C1 (G178D) from 0–250 ng did not affect the phosphorylation level of auto-phosphorylated GST-CIPK9. The presence or absence of each protein in the reaction mixture is shown as + or –, respectively.