To obtain information about the organization of the Ctcfl gene

There were no genes or ESTs which were differentially expressed at several time point. Validation of gene expression by real time RT PCR To verify the altered mRNA expression of the ECM genes COL3A1, SPARC, BGN and NID1 at 48 h of decid ualization, quantitative real time RT PCR was carried out utilising BAM7 the same RNA samples found in the microarray anal ysis, plus two additional RNA samples of each genotype, collected within the same way. In a significance degree of p 0. 05, there clearly was no statistical difference while in the abundance of 18S rRNA, COL3A1, BGN, SPARC or NID1 mRNA between IL11Ra,and IL11Ra womb. The difference in NID1 variety between IL11Ra, when only the products used in the microarray analysis were considered, and statistical significance was approached by IL11Ra uterus at r zero. Metastasis 0708. Validation of gene expression by immunohistochemistry Some genes found to be differentially expressed in womb in comparison with wild type at 48 h of decidualization were examined at the protein level by immunohisto chemistry using specific antibodies. Decidualizing and absolutely decidualized cells were recognized in adjoining,areas by immunostaining for the intermediate filament proteins desmin, well characterized being a marker for decid ual modification, Microarray data showing highly significant and reproduc ible increases in COL3A1 and BGN mRNA levels in IL11Ra uterus were reflected in enhanced staining inten sity for collagen III and biglycan in IL11Ra uterus com-pared to wild type, In each IL11Ra and wild type uterus, collagen III and biglycan were pri marily localized to the outer connective-tissue and smooth muscle cells of the myometrium, with diffuse staining in the cytoplasm of decidualized stromal Whilst the epithelial cells were negative, cells, Interstitial pockets main luminal and glandular epithelium and surrounding bloodstream also showed strong immunoreactivity for both protein. In the lack of IL 11R, stronger staining for collagen III was particularly noticeable while in the ECM luminal epithelium and fundamental around decidualizing stromal cells. SPARC in uterus in comparison to wild type, the localization of the proteins has not previously been identified in the decidu oma of wild type or NSC-66811 IL11Ra mice.