Identification of human myometrial target genes of the cAMP pathway: the role of cAMP-response element binding (CREB) and modulator (CREM and CREM₂) proteins

Supplementary figure

Supplementary figure

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• Supplementary figure - (b) Total RNA samples were isolated from stably transfected cell lines expressing CREB. Human U133 A array chips were hybridized with biotin-labelled cRNA representing the mRNA populations of cultured myometrial cells in the form of (i) non-transfected controls, (ii) empty-vector stably transfected controls and (iii) stably transfected experimental lines over-expressing CREB. Details are given in the Data analysis and first Results sections of the paper. Due to the comprehensive number of genes involved, the resultant list was split into two; a supplementary list (Fig. 3e) and a main list (shown here) including genes thought to be definitely, probably and possibly linked to myometrial activity. The most germane of these are also listed in Fig. 3a alongside notable results from the CREMa and CREMô2a experiments. (c and d) Total RNA samples were isolated from stably transfected cell lines expressing the (c) CREMá and (d) CREMô2á isoforms, and hybridized to Affymetrix Human U133 A microarray chips as described in the Materials and methods section and for supplementary Fig. 3b. Following scanning and data analysis, those genes found to be up- or down-regulated by 2-fold or greater in comparison to baseline expression were classified according to their function by GeneSpring software, and are listed here. (e) Total RNA samples were isolated from stably transfected cell lines expressing the CREB gene, and hybridized to Affymetrix Human U133 A microarray chips as described in the Materials and methods section and for supplementary Fig. 3b. Following scanning and data analysis, those genes found to be up- or down-regulated by 2-fold or greater in comparison to baseline expression were classified according to their function by GeneSpring software; those results classified as clones, unnamed sequences, hypothetical proteins, expressed sequence tags (ESTs) and open reading frames (ORFs) are listed here, along with annotated genes not yet thought to be directly linked with with myometrial activity. Genes thought to be definitely, probably and possibly linked to myometrial activity are listed in Fig. 3a and online supplementary Fig. 3b.