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. 2014 May 13;33(13):1454–1473. doi: 10.15252/embj.201488598

Figure 6. Blocking Wnt reduces in vivo tumor growth.

Figure 6

  1. Expression of dnLEF-1 or dnTCF-1Emut in SW480 cells results in smaller xenograft tumors. Images of tumors are shown with quantification of the tumor mass and volume at the time of harvest. Data include measurements of eight tumors for each condition. Error bars represent the SEM among eight replicates (***P-value < 0.001).
  2. Ki67 staining of paraffin-embedded sections from xenograft tumors shows fewer Ki67-positive cells with dnLEF-1/dnTCF-Emut expression. Data shown represent the average of counts from at least eight fields. Error bars represent the SEM among at least eight replicates (*P-value < 0.05).
  3. Western blot was performed on protein lysates prepared from freshly extracted xenograft tumors. Western blot analysis of endogenous PDK1 and its target, pyruvate dehydrogenase (pSer293-PDH), shows decreases when dnLEF-1 or dnTCF-1Emut tumors are expressed. cMYC expression shows variable levels of expression.
  4. Phasor plot representation for the color mapping of in vivo tumor FLIM analysis, fluorescence intensity (top image panels), FLIM color mapping (bottom panels), and scatterplot analysis are as described in Fig5 except that these analyses were performed on living, surgically exposed, yet still actively perfused, xenograft tumors. Both the free/bound NADH color mapping (bottom row) and scatterplot (average phasor position of individual cells within each tumor) show a shift in the phasor position toward bound NADH with dnLEF-1 expression and a return to free NADH with PDK1 overexpression (P < 0.0001 comparing dnLEF-1 to mock and comparing dnLEF-1+PDK1 to dnLEF-1). A minimum of three fields of view per tumor were analyzed. Data shown are from one mouse representative of eight replicates (additional tumor analysis in Supplementary Fig S7).

Source data are available online for this figure.