Figure 1
In vivo expression of COMP-Ang1 gene and protein.
(a) Detection of COMP-Ang1-specific mRNA in the corpus cavernosum from hypercholesterolemic mice. PCR was performed with primers specific for COMP-Ang1. RNA was extracted from the corpus cavernosum 3, 7, 14 and 21 days after intracavernous administration of ad-COMP-Ang1 (2 × 108 parts/20 μl). Densitometric data are presented as the relative ratio of COMP-Ang1 mRNA to β-actin mRNA. The relative ratio measured 3 days after injection of ad-COMP-Ang1 was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. M, marker. (b) Western blot analysis of the expression of COMP-Ang1 protein in cavernous tissues from hypercholesterolemic mice 1, 6 and 24 hours and 3, 7, 14 and 21 days after intracavernous injection of ad-COMP-Ang1 (left) or COMP-Ang1 protein (right). The relative ratio of COMP-Ang1 to β-actin measured 3 days after injection of ad-COMP-Ang1 or 1 hour after injection of COMP-Ang1 protein was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. (c) Anti-FLAG staining of cavernous tissue from hypercholesterolemic mice 7 days after intracavernous injection of ad-LacZ (2 × 108 parts/20 μl) or ad-COMP-Ang1 (2 × 108 parts/20 μl) and 1 hour after intracavernous injection of BSA (5.88 μg/20 μl) or COMP-Ang1 protein (5.88 μg/20 μl). 2nd Ab, secondary antibody control; arrowheads, endothelial cells; arrows, smooth muscle cells. Scale bar = 50 μm. (d) Immunohistochemical staining of cavernous tissue using antibodies to FLAG (red), PECAM-1 (CD31; green) and α-actin (green) in each group. Scale bar = 50 μm.
