Figure 5.

Drebrin inhibits SMC up-regulation of CD68 and Galectin-3. (A) Aortic SMCs from Dbn^flox/flox and SMC-Dbn^−/− mice were transduced with empty vector or Drebrin-encoding adenoviruses and then exposed to medium containing vehicle or solubilized cholesterol for 48 h (37°C). SMCs were then immunostained with IgG specific for CD68 or Drebrin (or isotype control IgG), as indicated, counter-stained for DNA, and imaged at 200× (original magnification). Scale bars = 50 μm. (B) For each SMC group, the number of CD68⁺ SMCs was divided by the total number of SMCs and multiplied by 100; the resulting percentages are plotted (along with means ± SE) for four experiments with four distinct pairs of Dbn^flox/flox and Dbn^−/− SMCs. P < 0.05 for comparisons of (i) Dbn^−/− vs. cognate Dbn^flox/flox SMCs (*); (ii) Drebrin adenovirus-transduced vs. cognate control adenovirus-transduced SMCs (§); (iii) cholesterol-treated vs. vehicle-treated SMCs (#) (two-way ANOVA with Sidak post hoc test). (C) Dbn^flox/flox (f/f) and Dbn^−/− SMCs were transduced and treated ± cholesterol just as in A. After the 48-h incubation ± cholesterol; however, SMCs were solubilized; protein extracts were immunoblotted serially for the indicated proteins. (D) The band intensities for Galectin-3 were normalized to cognate β-actin band densities, and these ratios were plotted (along with means ± SE) for three experiments with three distinct pairs of Dbn^flox/flox and Dbn^−/− SMCs. P < 0.05 for comparisons of (i) Dbn^−/− vs. cognate Dbn^flox/flox SMCs (*); (ii) cholesterol-treated vs. vehicle-treated SMCs (#); (iii) Drebrin adenovirus-transduced vs. cognate control adenovirus-transduced SMCs (§) (two-way ANOVA with Sidak post hoc test).