Fig. 8

Non-phosphorylated form of MYB3R3 represses the KNOLLE and CYCB1;2 promoters. a ChIP-PCR analysis of KNOLLE and CYCB1;2. Ten-day-old seedlings of myb3r3 carrying ProMYB3R3::MYB3R3-GFP (WT), ProMYB3R3::MYB3R3AAA-GFP (AAA) or ProMYB3R3::MYB3R3DDD-GFP (DDD) were treated with 2 μM zeocin for the indicated times, and used for ChIP assay. Chromatin bound to MYB3R3 was collected by immunoprecipitation with anti-GFP antibodies. Fold enrichment of each promoter region was determined by normalizing the recovery rate against that of ProMYB3R3::MYB3R3-GFP without zeocin treatment. Data are presented as mean ± SD of three biological replicates. Significant differences from ProMYB3R3::MYB3R3-GFP were determined by Student’s t-test: **P < 0.01; ***P < 0.001. b Quantitative RT-PCR analysis of KNOLLE and CYCB1;2. Five-day-old seedlings of WT, myb3r3 and myb3r3 carrying ProMYB3R3::MYB3R3-GFP, ProMYB3R3::MYB3R3AAA-GFP or ProMYB3R3::MYB3R3DDD-GFP were transferred to medium with 2 µM zeocin and grown for 24 h. Total RNA was isolated from roots and subjected to quantitative RT-PCR analysis. The expression levels were normalized to that of ACTIN2, and are indicated as relative values, with that for the time of transfer to zeocin-containing medium set to 1. Data are presented as mean ± SD of three biological replicates. Significant differences from the WT control were determined by Student’s t-test: ***P < 0.001