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Culture of mouse pancreatic beta cell line MIN6 cells were infected with GFP or ACREB adenovirus, and treated with cAMP agonist forskolin (FSK) for 1hr. RNA was extracted using the Qiagen RNeasy kit. Two controls and two forskolin treated samples from GFP or ACREB virus infected cells were analyzed.
Total RNA samples were labeled and hybridized by standard protocol to Affymetrix GeneChip Murine Genome U74 Version 2 Set MG-U74A array. Array data was analyzed using the dChip software with PM-MM models.
Guidelines for Choosing cAMP Induced Genes in this Data
The fold change (FC) value is positive for increased expression and negative for decrease expression. There is also a lower bound of fold change (LFC), which is a conservative estimation of the fold change with 90% confidence. For this data set, if a gene was induced by FSK in GFP infected samples (LFC>=1) and this induction was inhibited by ACREB ((FSK_ACREB vs FSK_GFP) LFC<=-1), we consider it as a good cAMP induced gene and therefore a potential CREB target.