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. Author manuscript; available in PMC: 2016 Jan 30.
Published in final edited form as: Oncogene. 2014 Oct 20;34(31):4069–4077. doi: 10.1038/onc.2014.335

Figure 2. A regulatory feedback loop between NR2E1 and CBX7.

Figure 2

(a) Scheme of the CBX7 gene showing the location of primers sets used for ChIP and probes used for DPA. (b) DNA pulldown analysis. DNA pulldown assay confirmed NR2E1 binding to a DNA motif upstream of CBX7 transcription start site. Lysates of HEK293T cells overexpressing NR2E1 wt or the DNA binding domain mutant NRE1 Δ40 were incubated with probe 4, harbouring a wt or mutated NR2E1 DNA binding motif. NR2E1 binding was assessed by immunoblot (IB). Black arrow, NR2E1 wt; blue arrow, NRE1 Δ40. (c) NR2E1 is a direct transcriptional regulator of CBX7. ChIP with FLAG antibody or control IgG were performed on CBX7 (left) or CDKN1A (right, as positive control) in IMR90 cells infected with FLAG-NR2E1 overexpressing vector or with an empty vector. (d) CBX6, CBX7 and CBX8 associate with NR2E1 promoter. ChIP-seq data in Hs68 HFs showing CBX6, CBX7 and CBX8 binding profiles on NR2E1. The location of primer sets used for ChIP on NR2E1 is also depicted. (e) ChIP in IMR90 cells confirming that CBX7 binds to NR2E1 promoter. (f) CBX7 regulate NR2E1 expression, as shown by qRT-PCR upon CBX7 overexpression (left) or knockdown (right) in IMR90 cells.