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. 2017 Apr 10;8:502. doi: 10.3389/fpls.2017.00502

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Copyright © 2017 Lyzenga, Sullivan, Liu and Stone.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Activated CIPK26 promotes the degradation of KEG in planta. (A) Cell-free degradation assay using protein extracts from 5-day-old OLexA:CIPK26^TD-YFP-HA, 35S:HA-KEG/keg-1 (line 1) and OLexA:CIPK26^KR-YFP-HA, 35S:HA-KEG/keg-1 (line 1) seedlings induced to express CIPK26 with 20 μM 17-β-estradiol. HA-KEG protein abundance was determined by WB using HA antibodies at the indicated time points. (B) Cycloheximide (CHX) chase assay using 5-day-old OLexA:CIPK26^TD-YFP-HA, 35S:HA-KEG/keg-1 (line 1) and OLexA:CIPK26 ^KR-YFP-HA, 35S:HA-KEG/keg-1 (line 1). Seedlings were incubated in liquid growth medium supplemented with or without 20 μM 17-β-estradiol to induce expression of CIPK26. Seedlings were then treated with CHX and samples collected at the indicated time points. The abundance of HA-KEG present at each time point was determined by WB with HA antibodies.