ABSTRACT: UMRR (Universal Mouse Reference RNA, Catalog #740100) was purchased from Stratagene and used as the starting material. Liver total RNA (Catalog #64042-1) was purchased from Clontech and used as the starting material. UMRR was labeled with Cyanine-3 (Green) while Liver was labeled with Cyanine-5 (Red). cRNA was made by the single round amplification method.
Project description:UMRR (Universal Mouse Reference RNA, Catalog #740100) was purchased from Stratagene and used as the starting material. Spleen total RNA (Catalog #64044-1) was purchased from Clontech and used as the starting material. UMRR was labeled with Cyanine-3 (Green) while spleen was labeled with Cyanine-5 (Red). cRNA was made by the single round amplification method.
Project description:Spleen total RNA (Catalog #64044-1) and liver total RNA (Catalog #64042-1) were purchased from Clontech and used as the starting material. Spleen was labeled with Cyanine-3 (Green) while liver was labeled with Cyanine-5 (Red). cRNA was made by the double round amplification method.
Project description:Spleen total RNA (Catalog #64044-1) and liver total RNA (Catalog #64042-1) were purchased from Clontech and used as the starting material. Spleen was labeled with Cyanine-3 (Green) while liver was labeled with Cyanine-5 (Red). cRNA was made by the single round amplification method.
Project description:Spleen total RNA (Catalog #64044-1) and liver total RNA (Catalog #64042-1) were purchased from Clontech and used as the starting material. Spleen was labeled with Cyanine-3 (Green) while liver was labeled with Cyanine-5 (Red). cRNA was made by the single round amplification method.
Project description:UMRR (Universal Mouse Reference RNA, Catalog #740100) was purchased from Stratagene and used as the starting material. Liver total RNA (Catalog #64042-1) was purchased from Clontech and used as the starting material. UMRR was labeled with Cyanine-3 (Green) while Liver was labeled with Cyanine-5 (Red). cRNA was made by the single round amplification method. Keywords: repeat sample
Project description:UMRR (Universal Mouse Reference RNA, Catalog #740100) was purchased from Stratagene and used as the starting material. Spleen total RNA (Catalog #64044-1) was purchased from Clontech and used as the starting material. UMRR was labeled with Cyanine-3 (Green) while spleen was labeled with Cyanine-5 (Red). cRNA was made by the single round amplification method. Keywords: repeat sample
Project description:Dickkopf 1 (DKK1), which is expressed at high mRNA levels by fibroblasts in the dermis of human skin on the palms and soles, inhibits the function and proliferation of melanocytes in the epidermis of those areas via the suppression of beta-catenin and microphthalmia-associated transcriptor factor (MITF). In this study, we investigated the effects of DKK1 on melanocyte gene expression profiles and on Wnt signaling pathways using DNA microarray technology. Paired cDNA samples, labeled by cyanine 3- and cyanine 5-dUTP incorporation (Qiagen, Valencia, CA) during reverse transcription (Qiagen), were hybridized simultaneously with one oligo-DNA chip (HS-Operon V2vB2.2p13) as per NCI in-house protocol (available at http://mach1.nci.nih.gov/). Two fluorescent intensities of the oligo-DNA chip were scanned using a microarray scanner (GenePix 4000A, Axon Instruments, Inc., Molecular Device Corp., Sunnyvale, CA).
Project description:We examined the major patterns of changes in gene expression in mouse splenic B cells in response to stimulation with 33 single ligands for 0.5, 1, 2, and 4 h. We found that ligands known to directly induce or costimulate proliferation, namely, anti-IgM (anti-Ig), anti-CD40 (CD40L), LPS, and, to a lesser extent, IL-4 and CpG-oligodeoxynucleotide (CpG), induced significant expression changes in a large number of genes. The remaining 28 single ligands produced changes in relatively few genes, even though they elicited measurable elevations in intracellular Ca2+ and cAMP concentration and/or protein phosphorylation, including cytokines, chemokines, and other ligands that interact with G protein-coupled receptors. A detailed comparison of gene expression responses to anti-Ig, CD40L, LPS, IL-4, and CpG indicates that while many genes had similar temporal patterns of change in expression in response to these ligands, subsets of genes showed unique expression patterns in response to IL-4, anti-Ig, and CD40L.
Project description:total RNA from mouse (male c57BL/6) spleen labeled with Cy3 vs total RNA from mouse (male c57BL/6) B cells treated with Neurokinin B (NEB) labeled with Cy5- time course with repeats
Project description:We identified differentially expressed genes in response to single and double ligand treatments (LPS, IFG, 2MA, LPS plus 2MA, and LPS plus IFG). The majority of the regulated transcripts responded additively to dual ligand treatment. However, a significant fraction responded in a non-additive fashion. Several cytokines showing non-additive transcriptional responses to dual ligand treatment also showed non-additive protein production/secretion responses in separate experiments. Many of the genes with non-additive responses to LPS plus 2MA showed enhanced responses and encoded pro-inflammatory proteins. On the other hand, LPS plus IFG appeared to induce both non-additive enhancement and non-additive attenuation of gene expression. The affected genes were associated with a variety of biological functions. These experiments reveal both dependent and independent regulatory pathways and further analyses may point out the specific nature of the regulatory interactions. Keywords = Lipopolysaccharide Keywords = Interferon-gamma Keywords = 2-Methyl-thio-ATP Keywords = Dual Ligand Effects Keywords = Gene Expression Keywords = RAW 267.4 Keywords = Macrophage