Figure 5

Effect of diabetes on nuclear estrogen receptor alpha (ERα) and androgen receptor (AR) in the rat vagina. Vaginal tissues from diabetic (Diab) or age-matched control (Con) rats were homogenized and the nuclear subcellular fraction was obtained by differential centrifugation. ERα was extracted from the nuclei by homogenization and incubation in high salt buffer. The vaginal tissue nuclear extract (200 μg total protein/lane) was then subjected to gel electrophoresis under denaturing conditions. Gels were blotted onto nitrocellulose membranes that were probed with antibody against ERα or AR. After incubation with the secondary antibody, membranes were developed using an enhanced chemiluminescence reaction and bands were visualized by exposure to film.