Ls b2138

Penis tissues were fixed by perfusion with 4% paraformaldehyde and then embedded in paraffin. The specimens were sliced at a thickness of 5 μm, deparaffinized, rehydrated, and subjected to histochemical analysis. Elastica van Gieson staining was performed according to the standard method. In brief, the specimens were stained with resorcin fuchsin staining solution (Muto Pure Chemicals, CO., LTD., Tokyo, Japan), hematoxylin staining solution, and van Gieson staining solution (Muto Pure Chemicals, CO., LTD., Tokyo, Japan). For immunohistochemistry, sections were incubated with a primary antibody reactive to VE-Cad (1:400; LS-B2138; LifeSpan BioSciences), SMA (1:200; ab18147; Abcam) or nNOS (1:400; ab76067; Abcam). Sections were then incubated with biotinylated secondary antibody prior to horseradish peroxidase-labeled streptavidin according to the manufacturer's instructions (DAKO, Cambridgeshire, UK).