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. 2014 Nov 17;211(12):2411–2423. doi: 10.1084/jem.20131857

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© 2014 Guiu et al.

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Figure 2. — Cdca7 is a direct Notch target gene through two predicted RBPj-binding sites. (A) Schematic representation of the promoter region of Cdca7 showing the array probe position (red circles), DNA (black dashed line), TSS (black arrow), putative RBPj-binding site (blue bars), and probe enrichment in log₂ ratio (gray bars). RBPj-binding sites were identified with the STORM and Genomatix software scanning from the −10- to 0-kb region of each target gene. (B) ChIP with anti-RBPj. (C) ChIP with anti-N1IC. (D) ChIP with anti-MAM1. In B–D, chromatin was extracted from E10.5 dissected AGMs (n = 2). (E and F) Luciferase activity from Cdca7 promoter (−511/50 bp) WT (E) and mutant RBP sites (F) in the presence of the specified transfected plasmids into HEK-293T cells (micrograms of transfected plasmid are indicated; n = 3). In the bottom panels, Western blot of the transfected proteins is shown. Error bars represent the SD. Significant differences are indicated by asterisks (*, P < 0.05; **, P < 0.01).