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. 2008 Apr 21;105(17):6392–6397. doi: 10.1073/pnas.0802047105

Fig. 3.

Fig. 3.

Activation of Notch can mimic hypoxia-induced EMT. (A) Phase contrast images of SKOV-3 cells grown on recombinant FC or FC-Jagged in the presence or absence of GSI in normoxia. The same number of cells was seeded in each experiment. (B) Western blot analysis of endogenous E-cadherin protein levels in SKOV-3 cells kept at normoxia or hypoxia after transfection of N1ICD. (C) Western blot analysis of E-cadherin expression in SKOV-3 cells after infection with N1ICD, Hes-1, Hey-1, or EGFP. (D) E-cadherin protein levels after cotransfection of N1ICD with dnCSL or dnMaml1 at normoxia. (E) 12XCSL-luciferase activation in 293T cells cocultured with wild-type (−) or Jagged1-expressing 293T 293T-J) cells transfected with dnMaml1, treated with hypoxia or transfected with HIF-1α(PP-A). (F) Coimmunoprecipitation analysis of HIF1α–N1ICD–Maml1 interaction. Notch was immunoprecipitated from 293T cells transfected with N1ICD, FLAG-Maml1, and FLAG-HIF-1α(PP-A) as indicated, and the presence of coimmunopecipitated Maml1 and HIF-1α was detected by Western blot analysis. The input control is shown below. Values are significant at **, P < 0.01 and *, P < 0.05 as indicated.