Increased Smad2 phosphorylation at Ser 465/467 in smLRP1−/−; LDLR−/− and smLRP1−/−; LDLR−/−; PDGFRβ F2/F2 aortas.
(A) Immunofluorescent staining of p-Smad2 (Ser 465/467, green) in aortas of atherosclerosis-free 1-month old mice. Scale bar, 20 µm. (B) Immunoblotting of aortic extracts for Smad2 phosphorylation at Ser 465/467. β-actin served as a loading control. (C) The density of p-Smad2 signals was normalized to the corresponding β-actin signals from the same blot and quantified using Image J software (NIH). Densitometric scanning from three independent experiments revealed a statistically significant average of 250%±13% in smLRP1−/− mice and 260%±29% in smLRP1−/−; PDGFRβ F2/F2 mice, compared with wild type controls (92%±17%). Relative ratio of p-Smad2 in mouse aortas from the indicated genotypes was plotted. Data are presented as mean±SD. * p<0.001, n = 3.