Figure 4.

Wnt3a protein inhibited H₂O₂‐induced apoptosis in VSMCs from young and old mice, whereas Wnt5a‐mediated rescue and WISP‐1 upregulation was lost with age. (a–d) Apoptosis was quantified in VSMCs isolated from young (a, c) and old (b, d) mice and stimulated with 100 μM H₂O₂, with or without 400 ng/ml recombinant Wnt3a (a, b) or Wnt5a protein (c, d), for 24 hr using CC3 immunofluorescence. The number of CC3 positive cells was counted and expressed as a percentage of total number of cells viewed. *p < 0.05 vs. control, $p < 0.05 vs. H₂O₂, repeated measures ANOVA and Student–Newman‐Keuls post hoc test, n = 3 for C and n = 5 for (a, b and d). (e, f) WISP‐1 mRNA was quantified by QPCR in VSMCs isolated from young (e) and old (f) mice and stimulated with 100 μM H₂O₂, with or without 400 ng/ml recombinant Wnt5a protein, for 4 hr. WISP‐1 mRNA levels were quantified from a standard curve. Results are shown as the fold change from control. Error bars represent mean ± SEM. *p < 0.05 vs. control, $p < 0.05 vs. H₂O₂, ANOVA and Student–Newman–Keuls post hoc test, n = 7 for (e) and n = 5 for (f)