Monitoring of Serial Presurgical and Postsurgical Changes in the Serum Proteome in a Series of Patients with Calcific Aortic Stenosis

Figure 1

Verification of iTRAQ ratios by Western blot analyses. (a) Western blot analyses with anti-ceruloplasmin (CP) antibody in presurgical (pre) and postsurgical (post) sera of patients P2 and P3 as well as in sera of control volunteer N1 at two different time points ( and ) were performed. (b) Western blot analyses with anti-fibronectin (FN) antibody in presurgical (pre) and postsurgical (post) sera of patients P3 and P5 were performed. Representative photos are shown. Some bands in the Western blot analysis with anti-FN antibody might be derived from proteolytic products of intact FN molecules. To confirm equal levels of proteins per lane, nonspecific proteins stained with Coomassie Brilliant Blue (CBB) are shown in the lowest panel. The intensity of each band that reacted with the corresponding antibody indicated by an arrow was measured. In the case of densitometric analysis of FN level, the intensity of the intact FN corresponding upper band was measured. Ratios of levels of ceruloplasmin in postsurgical sera to those in presurgical sera (1.0) in patient P2 (d) and patient P3 (e) were determined by band intensity. In addition, ratios of levels of fibronectin in postsurgical sera to those in presurgical sera in patient P3 (g) and patient P5 (h) were determined by band intensity. Similarly, ratios of levels of ceruloplasmin (c) and fibronectin (f) at to those at (1.0) in serum of control N1 were determined by band intensity. Means ± SE of triplicate experiments were calculated, and statistical analysis was performed using the paired t-test. . .

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)