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. Author manuscript; available in PMC: 2013 Dec 24.
Published in final edited form as: Oncogene. 2012 Dec 17;32(42):10.1038/onc.2012.530. doi: 10.1038/onc.2012.530

Figure 4.

Figure 4

Y654-β-catenin phosphorylation is required for hypoxia-induced EMT and β-catenin promotion of HIF1α transcriptional activity. (a) E-cadherin (green) and α-SMA (orange) staining of AECTs expressing Y654E or Y654F-β-catenin. Scale bar, 100 µm. (b) Immunoblots for collagen I, α-SMA, Twist, and HIF1α of AECTs with β-catenin deletion (Cre), cells expressing wt (W) and Y654E (E) or Y654F (F) mutant in normoxia or hypoxia for 56 h. (c) Conditioned medium from the cells above were concentrated for zymography with recombinant MMP-2 as control. (d) EMT markers from B and C were quantified using Image J and the mean value normalized to β-catenin level from 3 independent experiments were shown. ns, not significant. * indicates p<0.05 by t-test. (e) HRE reporter activity of 293 cells expressing W, E or F mutant β-catenin under normoxia or hypoxia for 16 hours. Renilla activity was used as internal control. Mean value of hypoxia/normoxia ratio from 4 independent experiments was shown. * indicates p<0.05 by t-test. 293 cell lysates were also blotted for Myc-tagged β-catenin and β-actin.