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. 2018 Jul 19;19(9):e45409. doi: 10.15252/embr.201745409

Figure 4. YY1 interacts with Fuz promoter in vivo and in vitro .

Figure 4

  • A
    Schematic representation of the human Fuz −1332/+574 promoter region. The Fuz +68/+574 was enlarged to show the detailed nucleotide sequence. A putative CpG island was found within Fuz +68/+574 and was defined as Fuz +117/+347CpG. “+1” is the transcriptional initiation site (arrow). The CpG island is highlighted in blue, and sequence of the YY1 binding site is underlined and highlighted in red.
  • B
    Chromatin immunoprecipitation assay demonstrated the binding between YY1 protein and Fuz +117/+347CpG DNA fragment. The blue bar indicates the Fuz +117/+347CpG sequence. However, Fuz −802/−612 (orange) represents a region that did not show interaction with YY1 protein. n = 3.
  • C, D
    (C) In vitro DNA binding assay demonstrated the binding between purified YY1 protein and purified Fuz +117/+347CpG DNA fragment. When a mutation (green) was introduced to the YY1 site (red) within the Fuz +117/+347CpG fragment, YY1 binding was reduced. (D) is the quantification of the intensity of the Fuz +117/+347CpG DNA bands as shown in (C). Error bars represent s.e.m., n = 3. Statistical analysis was performed using two‐tailed unpaired Student's t‐test. *P < 0.05.
  • E
    Mutating the YY1 binding site in the Fuz promoter increased its transcriptional activity in HEK293 cells. Error bars represent s.e.m., n = 5. Statistical analysis was performed using two‐tailed unpaired Student's t‐test. ***P < 0.001.
Data information: n represents the number of biological replicates. Only representative gels and blots are shown.Source data are available online for this figure.