Figure 2

Confirmation of iTRAQ ratio by Western blot analyses. Western blot analyses with anti-leucine-rich α-2-glycoprotein (referred to as LRG1) (A) and anti-fibronectin (FN) (B) antibodies in presurgical (pre) and postsurgical (post) sera of patient #AAA2 and patient #TAA4 were performed. Representative photos are shown. To confirm equal levels of proteins per lane, non-specific 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. Ratios of levels of LRG1 (C) and fibronectin (F) in postsurgical sera compared with those in presurgical sera (1.0) in #AAA2 were determined by band intensity. Ratios of levels of LRG1 (D) and fibronectin (G) in postsurgical sera compared with those in presurgical sera (1.0) in #TAA4 were also determined by band intensity. Furthermore, Western blot analyses with anti-LRG1 (A) and anti-FN (B) at two different time points (T1 and T2) in serum of a control volunteer #C3 were performed. Ratios of levels of LRG1 (E) and fibronectin (H) at T2 compared with those at T1 (1.0) in serum of the control volunteer were determined by band intensity. Means ± SE of quadruplicate experiments were calculated, and statistical analysis was performed using the paired t-test. ** p < 0.01. *p < 0.05.