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. 2018 May 15;37(11):e97499. doi: 10.15252/embj.201797499

Figure 4. SUP binds to YUC1/4 chromatin to achieve high levels of H3K27me3.

Figure 4

  1. Schematic drawing of the YUC1 genome structure showing regions amplified by primer sets used for ChIP analyses.
  2. ChIP binding assay of the SUP protein at the YUC1 genome. Relative enrichment of SUP‐GFP at the YUC1 locus, including the promoter and coding regions, was analyzed using the stage 4 floral buds from ap1 cal p35S::AP1‐GR sup‐5 pSUP::SUP‐GFP plants.
  3. Relative enrichment of H3K27me3 at the YUC1 locus is decreased in the stage 4 floral buds of ap1 cal p35S::AP1‐GR sup‐5 plants relative to ap1 cal p35S::AP1‐GR.
  4. Schematic drawing of the YUC4 gene structure showing regions amplified by primers used for ChIP analyses.
  5. ChIP binding assay of the SUP protein at the YUC4 gene. Relative enrichment of SUP‐GFP at the YUC4 locus, including the promoter and coding regions, analyzed using the stage 4 floral buds of ap1 cal p35S::AP1‐GR sup‐5 pSUP::SUP‐GFP.
  6. Relative enrichment of H3K27me3 is decreased at the YUC4 locus in stage 4 floral buds of ap1 cal p35S::AP1‐GR sup‐5 compared with that of the WT control. Error bars represent standard errors with three biological repeats.
  7. yuc1 and yuc4 mutant alleles partially rescue the stamen and carpel number defects of sup‐5, while yuc1 yuc4 is epistatic to sup‐5. The yuc4 mutation results in reduced floral organ size while yuc4 appears WT‐like. Both yuc1 and yuc4 single mutant have no defects in floral organ numbers. The yuc1 yuc4 double mutant shows strong floral defects, namely a decreased numbers of stamen and carpel together with a reduction in floral organ size. The sup‐5 yuc1 yuc4 triple mutant shows a yuc1 yuc4‐like floral morphology. A typical flower is shown for WT, yuc1, yuc4, sup‐5, sup‐5 yuc1, sup‐5 yuc4, yuc1 yuc4, and sup‐5 yuc1 yuc4, respectively. For yuc1 yuc4 and sup‐5 yuc1 yuc4, the top view and side view are shown for the same flower. Scale bars: 1 mm.
  8. The statistical analysis showed a reduction in total stamens and carpels in sup‐5 yuc1 (8.60 ± 1.33 for stamen, 2.13 ± 0.34 for carpel, n = 30), sup‐5 yuc4 (10.93 ± 1.74 for stamen, 2.4 ± 0.50 for carpel, n = 30) once compared with sup‐5 (14.00 ± 1.68 for stamen, 2.7 ± 0.75 for carpel, n = 30). *P < 0.05 based on Student's t‐test. There is no significant difference between yuc1 yuc4 (3.8 ± 0.77 for stamen, 1.25 ± 0.44 for carpel, n = 20) and sup‐5 yuc‐1 yuc4 (4.05 ± 0.70 for stamen, 1.05 ± 0.23 for carpel, n = 19) in floral organ number and floral organ size, P > 0.05 with a Student's t‐test.
Data information: In (B, C, E and F), error bars indicate s.e.m. of three biological replicates. In (H), error bars indicate s.d.