ategories that were enriched in the control groups, five growth r

ategories that were enriched in the control groups, five growth related GO sets, one epigenetics GO set, and one angiogenesis 17-DMAG hsp90 GO set. No gene set was enriched in the alcohol treated group. An example of gene enrich ment analysis is shown in Figure 4 for GO,0040007, Growth. This gene set contained 75 genes. The GSEA p values for this enrichment score were 0. 010 in Experi ment 1 and 0. 005 in Experiment 2. The growth related genes represented the largest group of affected genes. There were 5 GO sets of growth associated genes. Many of these genes, identi fied by GSEA in both experiments, were also identified in Experiment 2 at the single gene level, e. g. the Growth Gap43, Crim1, Tgfb3, Nov, Socs2, and Wrn were signifi cantly reduced in Experiment 2, and Igfbp7, Emp3, Bmp4, Bmp6, Inhbb, Wig1, and Cish were reduced but did not reach the criteria for significance.

The additional growth genes in Epidermal growth factor receptor signaling pathway GO group appear to be reduced to a greater extent in ALC NTO than in ALC NTC. b. Inhibitors,Modulators,Libraries Stem Cell Related Gene Sets Three gene sets were enriched in the control embryos compared to the combined alcohol treated embryos, TGF Beta activin responsive genes, extracellular matrix molecules, and ECM protease Inhibitors,Modulators,Libraries inhibitors. Three gene sets were down regulated in the ALC NTC subgroup, other related growth factors, other regulators of cell differentiation, and ECM protease inhibitors. Two gene sets were down regulated in the ALC NTO group, other related growth factor and other ECM molecules. There were no significant gene sets in compari sons between ALC NTC and ALC NTO.

No gene set was enriched in any alcohol treated group. Validation by Quantitative RT PCR Quantitative RT PCR was used to verify some of Inhibitors,Modulators,Libraries the genes that were significantly affected Inhibitors,Modulators,Libraries by alcohol, including a sample Carfilzomib of genes from the functional gene sets for neural specification and trophic factors identified in GSEA. These studies used independent embryos subjected to identical ethanol exposure. The qRT PCR verified that all 11 down regulated neural specification genes and neurotrophic growth factor genes tested dif fered in the same direction. One gene that did not differ in the microarray experiments was also tested and the lack of difference was confirmed. Discussion 1.

Developmental Deficits and Correlation with Gene Expression Profiles The abnormal embryonic development resulting from the alcohol treatment at this specific sellekchem stage of develop ment was consistent with our pre vious report and those of others. Two different facets of abnormal development could be iden tified, growth delay and frank teratogenesis. Delays in growth were also evident by the significant reductions in the total RNA per embryo and in the delayed morpholo gical staging. The affected structures were derived from each of the three germ layers, i. e. neural tube and brain vesicles, somites and cardio vascular system, and involved a wide range of tissues and organs. Alterations in all of these

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