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. 2016 Apr 8;25(11):848–860. doi: 10.1089/scd.2015.0362

FIG. 4.

FIG. 4.

Low oxygen tension stimulates the canonical Wnt pathway. Low oxygen increases (A) nuclear accumulation of β-catenin. (B) β-Catenin/TCF4 interactions: immunoprecipitation of 200 μg protein samples with TCF4, which were size separated and probed with β-catenin Abs in MIAMI cells grown at 3% oxygen (lane 1) and 21% oxygen (lane 2). Isotypic negative controls (normal rabbit serum); 3% samples (lane 3) and 21% (lane 4) Coomassie blue stain (bottom panel) as loading control. (C) CyclinD1 cytoplasmic (lanes 1–2) and nuclear proteins (lanes 3–4) at 3% and 21% oxygen. Coomassie stain is loading control. (D) Changes in the levels of cytoplasmic and nuclear β-catenin in response to oxygen and Wnt3a stimulation. Changes in the levels of cytoplasmic and nuclear Oct4 in response to oxygen and Wnt3a stimulation. (E) Low oxygen fails to stimulate Wnt3a secretion: ELISA from MIAMI cell-conditioned media taken from cells grown for 7 days at 3% and 21% oxygen. A media-only sample was run to determine baseline levels of Wnt3a. (F) Exogenous Wnt3a increased the cell number at low and high oxygen tension: total cell numbers of cells seeded at 100 cells/cm2 and grown at 3% or 21% oxygen with or without rmWnt3a treatment for 7 days. **P ≤ 0.01 and ***P ≤ 0.001.