Figure 2. Mechanism of the angiogenesis in the TMA.

Immunohistochemical staining of CD34, vWF, KDR and p-KDR showed that positive cells labeled brown of fresh lung tissue (0 day) and cultured 9 days lung tissue from the TMA. Representative immunostaining pictures are presented (a₁–d₃). Magnification: 200×. (e) Change of VEGF secretion in culture system supernatant. The supernatant was collected every other day and used to quantitative the VEGF. The expression of VEGF in supernatant was evidently increased along with the time went on. Values are expressed as means ± SEM. (n = 2). *P < 0.05, **P < 0.01 versus the 5th day group. (f) Effect of VEGF on the TMA. VEGF was added to the culture system, vessels grew quicker than ones in control group. Values are expressed as means ± SEM. (n = 4). (g) Quantification of protein expression from 4 sections in each group. Data were expressed as mean values ± SEM. *P < 0.05, **P < 0.01 versus the 0 day group. (h) Western blot analysis of CD34, vWF, KDR and p-KDR expression in fresh lung tissue (0 day) and cultured lung tissues in the TMA (cultured 9 days). Gels were run under the same experimental conditions and GAPDH is shown as a control. The cropped gels images are shown in and the full-length blots/gels are presented in Supplementary Figure 1. (i) Quantification of CD34 and vWF protein expression. Data were expressed as mean values ± SEM. *P < 0.05, **P < 0.01 versus the 0 day group. (j) Quantification of p-KDR protein expression. Data were expressed as mean values ± SEM. *P < 0.05 versus the 0 day group.