Figure 5.

Constitutive activation of IKKβ in vascular smooth muscle cells (VSMCs) inhibits β‐catenin–Runt‐related transcription factor 2 (Runx2) signaling. A, Representative Western blots and densitometric analysis of active and total β‐catenin/GAPDH and Runx2/GAPDH expression in a whole cell lysate of cultured VSMCs isolated from wild‐type (WT) and kinase‐active IKKβ (KA) littermate mice. Bars represent the mean±SD (t test, n=4). B, Representative Western blots and densitometric analysis of Runx2 expression in nuclear and cytoplasmic extractions normalized to Tata‐binding protein (TBP) or GAPDH, respectively. Bars represent the mean±SD (t test, n=3). C, Representative image of 3 independent ubiquitination assays in which β‐catenin was precipitated and blotted with antiubiquitin. D, Representative images of immunostaining of the aorta section from WT, IKKβ knockout (KO), and KA mice 2 weeks after saline (sham) or CaCl₂ treatment with antiactive β‐catenin and Runx2 antibodies. Bar=20 μm. *P<0.05, ***P<0.001.