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. 2018 Mar 22;30(5):1062–1076. doi: 10.1105/tpc.17.00845

Figure 6.

Figure 6.

PRU1 Mediates WRKY6 Degradation Under Low-Pi Stress.

(A) and (B) Cell-free degradation assay. Seven-day-old pru1 mutant and wild-type seedlings were transferred to MS medium, LP medium, or LP medium containing 10 μM MG132 for 3 d, and then the roots were harvested and protein was extracted. The root protein extracts were incubated with recombinant WRKY6-His (A) or its deletion derivatives (WRKY6N-His, WRKY6M-His, or WRKY6C-His) (B) for the indicated periods, and the abundance of WRKY6 or its deletion derivatives was determined by immunoblotting with anti-His antibody. Recombinant WRKY6-His or its derivatives in incubation buffer (without root proteins) were used as controls.

(C) Immunoblot analysis of WRKY6 in the pru1 mutant, complementation lines (COM5 and COM7) and wild-type plants. Seven-day-old seedlings were transferred to MS medium, LP medium, or LP medium containing 10 μM MG132, and then the roots were harvested at the indicated time points and protein was extracted. The abundance of WRKY6 was analyzed by immunoblotting using an anti-WRKY6 antibody. ACTIN was used as the loading control.