Figure 9.

AtMIF2/SlIMA Bind WUS/SlWUS Locus to Repress Their Expression via Histone Deacetylation.

(A) Schematic representation of the WUS and SlWUS loci (5′ and 3′UTR are in gray, introns in white, and exons in black) with the different targeted regions (P1 to P8).

(B) ChIP assay at the WUS locus using HA- or IgG-antibodies in Col-0, knu, Pro35S:AtMIF2-3HA in Col-0, and Pro35S:AtMIF2-3HA in knu plants. The y axis shows enrichment relative to input using IgG as a control. Error bars represent sd of three biological replicates, and asterisks indicate significant differences from the control (Col-0) using two-tailed t test (*P < 0.05 and **P < 0.01).

(C) ChIP assay at the SlWUS locus using GFP or IgG antibodies in wild-type and Pro35S:IMA-YFP plants. The y axis shows relative enrichment to input using IgG as a control. Error bars represent sd of three biological replicates, and asterisks indicate significant differences from the control (Col-0) using two-tailed t test (*P < 0.05 and **P < 0.01).

(D) DamID ratios at different regions of the WUS locus prior (NI) and after 24 h of ethanol induction (I) in lines expressing Arabidopsis MIF2 or KNU fused to DAM (Dam-AtMIF2 and Dam-KNU, respectively).

(E) Expression analysis of WUS in Col-0, Pro35S:AtMIF2, and hda19 floral buds (stages 1–12) treated or not with 0.5 µM TSA, using qRT-PCR. Error bars represent sd of three biological replicates and asterisks indicate significant differences from the control (Col-0) using two-tailed t test (*P < 0.05 and **P < 0.01).

(F) Expression analysis of WUS using GUS staining of ProWUS:GUS Pro35S:AtMIF2 double transgenic plants treated (upper panel) or not (lower panel) with 0.5 µM of TSA. Bars = 10µm.