Figure 5.

Wnt5a‐mediated activation of CREB, but not β‐catenin, was impaired by age. (a, b) β‐catenin nuclear translocation was quantified in VSMCs isolated from young (a) and old (b) mice and stimulated with 400 ng/ml recombinant Wnt5a protein for 30 min by immunofluorescence. The number of cells with β‐catenin nuclear translocation, defined as perinuclear β‐catenin staining, was counted and expressed as a percentage of the total number of cells viewed. Error bars represent mean ± SEM. *p < 0.05 vs. control, paired Student’s t test, n = 6 for (a) and n = 5 for (b). (c–f) Phosphorylated CREB (ser133) was detected by western blotting of VSMCs from young (c) and old (d) mice and stimulated with 400 ng/ml recombinant Wnt5a protein for 10 min. Levels of phosphorylated CREB (ser133) were normalized to β‐actin. *p < 0.05 vs. control, paired Student’s t test, n = 3. Representative western blots are shown (e, young and f, old). (g, h) HAS1 mRNA was quantified by QPCR in VSMCs isolated from young (g) and old (h) mice and stimulated with 400 ng/ml recombinant Wnt5a protein for 2 hr. HAS1 mRNA levels were normalized to 36B4 mRNA levels. Results are shown as the fold change from control. *p < 0.05 vs. control, one‐sample t test, n = 3 young and n = 4 old