Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from Mus musculus tissues (Heart, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:We identified genes expressed in mouse liver that are regulated by Cux2, a highly female-specific liver transcription factor whose expression is regulated by sex-dependent plasma GH patterns. Using siRNA to knockdown Cux2 expression in female liver, we show that female specific genes are predominantly repressed by Cux2 knockdown. In contrast, similar numbers of male-biased genes are repressed as are induced by Cux2 knockdown. A scrambled, non-specific siRNA was used as a control. (Published in Molec Cell Biology, TL Conforto et al, 2012) Liver RNA isolated from the following 3 groups of mice was used in the present study: (1) 8 wk old female mice treated with non-specific siRNA control (n = 13; 6 or 7 per each pool); (2) 8 wk old female mice treated with Cux2 siRNA and euthanized 5 days later (n = 5; 2 or 3 per each pool); (3) 8 wk old female mice treated with Cux2 siRNA and euthanized 8 days later (n = 4; 2 per each pool). These RNA pools were used in two separate sets of competitive hybridization experiments: 1) 8 wk non-specific siRNA treated vs. 8 wk Cux2 siRNA treated for 5 days; 2) 8 wk non-specific siRNA treated vs. 8 wk Cux2 siRNA treated for 8 days. Fluorescent labeling of RNA and hybridization of the Alexa 555-labeled (green) and Alexa 647-labeled (red) RNA samples to Agilent Mouse Gene Expression 4x44k v1 microarrays (Agilent Technology, Palo Alto, CA; catalog # G4122F-014868) were carried out, with dye swapping for each of the two hybridization experiments to eliminate dye bias. Two microarrays, one for each mixed cDNA sample, were hybridized for each of the two fluorescent reverse pairs, giving a total of 4 microarrays.
