Project description:This SuperSeries is composed of the following subset Series:; GSE9973: Half-life determination for human B-cells (BL41); GSE9975: newly transcribed RNA (nt-RNA) for IFN alpha and gamma time course; GSE9977: Expression data from NIH-3T3 cells treated with mock, 100 U/ml IFN alpha or 100 U/ml gamma for 1or 3h; GSE10011: Expression data from NIH-3T3 cells used for half-life determination Experiment Overall Design: Refer to individual Series
Project description:Data from tc-, nt- and p-RNA as well as 1 and 2h of actinomycin-D treatment (5µg/ml) of NIH-3T3 cells used to determine half-lives. RNA was labeled for 15, 30 or 60 minutes with 4-thiouridine. After preparation of tc-RNA, thiol-labeled RNA was biotinylated using biot-HPDP and subsequently tc-RNA was separated into nt- and p-RNA using streptavidin coated magnetic beads. All three fractions were used for microarray analysis. For actinomycin-D experiments only tc-RNA was used prepared from cell before and 1 an 2h after addition of act-D. We used microarrays to analyze the effects of 1 and 3h of IFNalpha and gamma treatment in total cellular RNA Keywords: determination of RNA half-lives in NIH-3T3