SREBP1c and mock virus expression profiles in AG01518 human fibroblasts

Sterol-regulatory element binding proteins (SREBP) control the expression of most genes involved in fatty acid and cholesterol metabolism. In this report, using microarrays, we showed that SREBP1 also induced genes unrelated to lipid biosynthesis, such as heme oxygenase 1 (HMOX1), the phosphatidylinositol-3 kinase regulatory subunit p55-gamma, synaptic vesicle glycoprotein 2A (SV2A) and COTE1 (C1orf2). These genes were regulated by infection with adenoviruses encoding active SREBP1, sterols, statins and growth factors, as one would expect from SREBP1 target genes Keywords: treatment comparaison Subconfluent AG01518 fibroblasts were infected with adenoviruses encoding SREBP1c or mock virus at 60 virus per cell for 24 h, which was shown to be optimal. Total RNA was isolated using the RNeasy kit (Qiagen) and used for microarray hybridizations as described (8). Briefly, total RNA (40 µg) from cells infected with SREBP1c or mock viruses were labelled in reverse transcription reactions (Superscript II kit, Invitrogen) with dCTP-Cy5 and dCTP-Cy3, respectively (Amersham). Purified cDNA probes labelled with Cy3 and Cy5 were mixed per pair, and hybridized to cDNA microarray chips (hver1.2.1) from the Sanger Institute/LICR/CRUK Consortium (see http://www.sanger.ac.uk/Projects/Microarrays/ for details and hybridization protocols). We performed three independent hybridizations, including one dye-swap control