Effect of DES on tissue specific expression of chicken <i>AHCYL1</i>.

<p>Both RT-PCR [A and B] and q-PCR [C and D] analyses were performed using cDNA templates from DES-treated and untreated oviducts. These experiments were conducted in triplicate and normalized to control <i>GAPDH</i> expression. [E] <i>In situ</i> hybridization analyses revealed cell-specific expression of <i>AHCYL1</i> mRNA in oviducts of DES-treated and untreated chicks. Cross-sections of the four segments of chicken oviduct (infundibulum, magnum, isthmus, and shell gland) treated with DES or vehicle were hybridized with antisense or sense chicken <i>AHCYL1</i> cRNA probes. [F] Immunoreactive AHCYL1 protein in oviducts of DES-treated and untreated chicks. For the IgG control, normal goat IgG was substituted for the primary antibody. Sections were not counterstained. See <i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049204#s4" target="_blank">Materials and Methods</a></i> for complete description. Legend: Untreated oviduct, non-treated whole oviduct; DES Treatment, DES treated whole oviduct; LE, luminal epithelium; GE, glandular epithelium; <i>Scale bar</i> represents 100 µm. The asterisks denote statistically significant differences (***<i>P</i><0.001 and **<i>P</i><0.01).