Figure 5.

AG Binds the WUS locus and Represses WUS Expression Directly.

(A) ChIP using anti-AG antibodies to determine AG occupancy at WUS. The null allele ag-1 and the eIF4A1 locus both served as negative controls. AP3, a known direct target of AG (Gómez-Mena et al., 2005), served as a positive control.

(B) Real-time RT-PCR to determine WUS transcript levels in 35S:AG-GR ag-1 inflorescences containing stage 8 and younger flowers. Inflorescences were treated with DMSO, DEX, CHX, or CHX plus DEX. Two hours later, the inflorescences were dissected to remove old flowers and harvested for RNA extraction and RT-PCR. Four biological replicates were performed for the DMSO/DEX experiment, and five were performed for the CHX/DEX experiment. Error bars represent sd, which were calculated from these biological repeats. The calculated P values for both experiments were 0.011.

(C) Real-time RT-PCR to measure WUS transcript levels in 35S:AG-GR ag-1 clf-47 inflorescences. Chemical treatments and RNA isolation were as in (B).