Journal: PLoS ONE

Article Title: CDC2 Mediates Progestin Initiated Endometrial Stromal Cell Proliferation: A PR Signaling to Gene Expression Independently of Its Binding to Chromatin

doi: 10.1371/journal.pone.0097311

Figure Lengend Snippet: A) The categories of over-represented Gene Ontology terms (GO terms) are shown by their decreasing p-values. The categories were identified by GOTM (Gene Ontology Tree Machine) software over the statistical regulated genes as indicated in supplementary Materials and Methods. Up-regulated genes percentages are shown in red, and down-regulated percentages are shown in green. B) Venn diagram shows the set of TF and CCR regulated by progestin. A GOTM search of cellular proliferation and cell cycle GO terms identified 23 genes, named cell cycle regulators (CCR) in the Figure. Seven of them were also categorized as transcription factors (TF) present in A. Up-regulated genes are shown in red, and down-regulated are shown in green. C) The signaling pathways associated to the differential gene expression pattern are shown by their decreasing p-value. Pathways identified by Pathway Express Software containing at least four progestin-dependent regulated genes included in a given Signalling Pathway (SP), with a p-value≤0.05. The percentage of up-regulated genes within a given signalling pathway is shown in red, and down-regulated genes are shown in green. Statistical details are described in M&M. D) The table shows individual fold changes of three independent biological samples (1,2,3) and one dye swap data set (1DS) analyzed by microarray, and the mean fold change of all 4 values (Media). Fold changes over vehicle treated cell values were calculated as described in SI M&M. E) q- real time PCR validation for JunD , Usf1 , Crebbp , Gfi1 , Cyr61 , Pten and Cdkn1b mRNA relative to β- Actin . The figure shows media ± SEM from three to six independent experiments. *P<0.05, **P<0.01, ***P<0.001 v. vehicle treated cells. F) sq-PCR validation for transcription factors (TF) JunD, Usf1 , transcription factors and cell cycle regulators (TF+CCR) Crebbp, Gfi1, Cyr61 , and cell cycle regulators (CCR) Pten, Cdkn1b and β-Actin .

Article Snippet: Isolated RNA was hybridized to an oligo microarray (60 mer) from Agilent (G4130). cDNA was synthesized according to manufacturer’s instructions (Agilent).

Techniques: Software, Expressing, Microarray, Real-time Polymerase Chain Reaction