Fig. 4.

In vitro degradation assay of MYB3R3. Recombinant His-MYB3R3 protein was incubated with or without root extracts from 7-day-old WT seedlings, and His-MYB3R3 was immunoblotted with anti-His antibodies. a Protein mixtures were incubated in the presence or absence of 50 µM MG132, 25 µM roscovitine and/or 10 mM ATP for the indicated times. b Total protein was extracted from 7-day-old WT roots treated with or without 2 μM zeocin for 24 h. Protein mixtures were incubated in the presence or absence of 50 µM MG132 for the indicated times. c Recombinant His-MYB3R3 or His-MYB3R3AAA proteins were incubated with root extracts in the presence or absence of 50 µM MG132 for the indicated times. Relative levels of His-MYB3R3 and His-MYB3R3AAA are expressed as the fold change, normalized with respect to the band of recombinant proteins visualized by CBB staining