Figure 4

In situ hybridization and immunohistochemical analysis of COX-2 expression in the ovine endometrium (Study Two). Ewes were ovariectomized on Day 5 of the estrous cycle, treated daily with progesterone, and infused from Days 11 to 15 with either control (CX) proteins or recombinant ovine IFNτ (IFN). On Day 16, ewes were hysterectomized. Cross-sections of the ovine uterus were hybridized with radiolabeled antisense or sense bovine COX-2 cRNA probe. Hybridized sections were digested with ribonuclease A, and protected transcripts were visualized by liquid emulsion autoradiography. Developed slides were counterstained lightly with hematoxylin, and photomicrographs were taken under bright-field or dark-field illumination (left). Immunoreactive COX-2 protein was detected using rabbit anti-human COX-2 polyclonal IgG (right). The negative IgG control was performed by substituting irrelevant rabbit IgG for primary antibodies. The white arrow denotes areas of specific COX-2 mRNA or immunoreactive protein expression. Legend: CX, control; IFN, interferon tau; LE, lumenal epithelium; S, stroma; sGE, superficial ductal glandular epithelium. Bar = 20 μm.