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Fasting/Refed: eight-week old male C57BL6/J mouse was fasted for 20 hr to induce glucagon-mediated cAMP signaling in hepatocytes (fasting sample). Control group (refed sample) was fasted for 16 hr then given food for 4 hr. Liver RNA was extracted using Trizol reagent and purified using the Qiagen RNeasy kit.
In parallel, mouse primary hepatocyte culture was treated with cAMP agonist forskolin (FSK) for 1hr or 4hr and total RNA was extracted using the Qiagen RNeasy kit.
Total RNA samples were labeled and hybridized by standard protocol to Affymetrix GeneChip Mouse Genome 430 2.0 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 fasting (LFC>=1) in liver and was also induced by FSK in hepatocyte in at least one of the two time points (LFC>=1 at 1hr or 4hr), then we consider it as a good cAMP induced gene in liver and therefore a potential CREB target.