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. 2016 Aug 24;6:31988. doi: 10.1038/srep31988

Figure 6. The stability of the CjWRKY1 protein in C. japonica 156-S protoplasts.

Figure 6

(a) The different stabilities of WT and mutant CjWRKY1 proteins. CjWRKY1 proteins were detected using CjWRKY1-specific antibodies. (b) A proteasome inhibitor, MG132 (50 μM), stabilized the CjWRKY1 proteins for 48 h. (c) Synergistic effects of MG132, protease inhibitors, and protein phosphatase inhibitors on the accumulation of CjWRKY1 proteins. Cj156-S protoplasts were treated with 50 μM MG132, 1 mg/ml complete EDTA-free protease inhibitor cocktail, and/or 5 mg/ml phosSTOP phosphatase inhibitor cocktail. An acrylamide gel containing 50 μM Phos-tag was used to detect phosphorylated WRKY1. Arrows indicate the shifted bands corresponding to phosphorylated CjWRKY1. DMSO (0.1%) was used for mock treatments in (b,c). All experiments were repeated at least three times with similar results.