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. 2018 Sep 14;37(20):e99395. doi: 10.15252/embj.201899395

Figure 1. Identification of BCL9 as a novel mitotic Wnt signalling regulator to stabilise proteins and promote oncogenesis.

Figure 1

  • A
    Normalised Wnt/STOP signalling activity by quantification of the protein expression of two Wnt/STOP target proteins, β‐catenin and c‐Myc, in mitotic (M) and asynchronised cells (A) after silencing 47 Wnt signalling regulators.
  • B
    ELISA validation of selected Wnt/STOP signalling regulators (n = 3).
  • C, D
    (C) Immunoblotting and (D) quantification of the stability of four STOP target proteins in both G1S and natural mitotic cells treated with BCL9 siGenome pools and CHX (n = 3).
  • E
    Representative pictures (left panel) and the normalised biosensor intensity based on live‐cell imaging of the cells with a GSK3‐GFP biosensor following different treatments before and after metaphase (n = 10). Scale bars represent 10 μm.
  • F
    Venn diagram analysis of the number of Axin1, APC, and Wnt3a signalling and/or BCL9‐regulated transcriptional targets.
  • G
    Myc and β‐catenin target gene enrichment analysis based on the transcriptome targets regulated by BCL9.
  • H
    The representative xenograft tumours (top panel) and tumour growth analysis (bottom panel) of scramble and BCL9 stable knockdown cells. LF: left flank of mouse, RF: right flank. n = 6 for shS1 vs. shB1, n = 5 for shS1 vs. shB2.
  • I
    Immunohistochemical staining of the xenograft; the red arrows indicate the representative mitotic cells. Scale bars represent 200 μm.
Data information: Significance was measured by two‐tailed unpaired t‐test, *P < 0.05, **P < 0.01, ***P < 0.001. All data are the mean ± SD. See also Fig EV1 and Tables EV1, EV2 and EV3.Source data are available online for this figure.