Figure 3 from BRCA Status Dictates Wnt Responsiveness in Epithelial Ovarian Cancer

Characterization of the Wnt signaling pathway in isogenic mouse ovarian cancer cells upon loss of Brca1 or Brca2. Cellular fractionation was performed to separate cytoplasmic (A) and nuclear (B) fractions in control no-treatment cells or cells treated with 100 ng/mL Wnt3A for 8 hours; ILF3 and Cox-IV were used as integrity controls for nuclear and cytoplasmic fractions, respectively. C, Levels and cellular location of β-catenin were determined by immunofluorescence for β-catenin (green). DAPI nuclear staining is shown in blue. Images show both green and DAPI channels; white arrows show increased green staining in the nucleus after Wnt3A treatment in ID8Trp53−/−; the red arrow shows increased green staining in the cytoplasm after Wnt3A treatment in ID8Trp53−/−;Brca1−/−; yellow arrows show membranal staining in ID8Trp53−/−;Brca2−/−; images shown are specific foci in the cultures. D, Levels of canonical Wnt/β-catenin gene targets were determined by Western blot analysis using whole-cell lysates. The red box shows truncated forms of Tcf1/7. DAPI, 4′,6′-diamidino-2-phenylindole.