Dissecting Stages of the Notch signaling pathway during Kidney Development by Oligonucleotide Array Sequence Analysis
ABSTRACT: Samples E12/E13/E14/E16/E18: We aims to screen out different gene expression profile in Embryo kidney on different gestation stages of the Notch signaling pathway Results from the various study components can help to screening important candidate genes during embryonic kidney development. Keywords: Embryo kidney, Development, Notch signaling pathway, Oligonucleotide Array Sequence Analysis The Oligo GEArray Assay comprises various components: RNA isolation,Assesing RNA yield and quality,cRNA labeling and synthesis,Hybridization,Chemiluminescent detection and Image acquisition and data analysis. Samples E12: This study has been accomplished with Embryo kidney on gestation 12, 3 techinical replicates. Samples E13: This study has been accomplished with Embryo kidney on gestation 13, 3 techinical replicates. Samples E14: This study has been accomplished with Embryo kidney on gestation 14, 2 techinical replicates. Samples E16: This study has been accomplished with 1Embryo kidney on gestation 16, 2 techinical replicates. Samples E18: This study has been accomplished with Embryo kidney on gestation 18, 2 techinical replicates.
Project description:Samples E12/E13/E14/E16/E18: We aims to screen out different gene expression profile in Embryo kidney on different gestation stages of the Notch signaling pathway Results from the various study components can help to screening important candidate genes during embryonic kidney development. Keywords: Embryo kidney, Development, Notch signaling pathway, Oligonucleotide Array Sequence Analysis Overall design: The Oligo GEArray Assay comprises various components: RNA isolation,Assesing RNA yield and quality,cRNA labeling and synthesis,Hybridization,Chemiluminescent detection and Image acquisition and data analysis. Samples E12: This study has been accomplished with Embryo kidney on gestation 12, 3 techinical replicates. Samples E13: This study has been accomplished with Embryo kidney on gestation 13, 3 techinical replicates. Samples E14: This study has been accomplished with Embryo kidney on gestation 14, 2 techinical replicates. Samples E16: This study has been accomplished with 1Embryo kidney on gestation 16, 2 techinical replicates. Samples E18: This study has been accomplished with Embryo kidney on gestation 18, 2 techinical replicates.
Project description:BMSC gene expression profiling was completed to examine the baseline neurotrophin and receptor gene expression in BMSC and to evaulate changes in gene expression in response to NGF or BDNF stimulation In the study, primary BMSC were treated with recombinant human NGF or BDNF for 24 hours. Following treatment, RNA isolated from the BMSC were analyzed using the Human Neurotrophin and Receptor Gene Array HS-018
Project description:Divergence has occured between the B10.BR-H2k H2-T18a/SgSnJJrep and B10.BR-H2k H2-T18a/SgSnJ (drifted) mouse strains, resulting in altered antigenic recognition and differential bone marrow engraftment capability. The microarray data demonstrate that the transcriptional profile of genes associated with hematopoiesis differs between lineage negative (as a marker for hematopoietic stem cells) bone marrow cells isolated from the B10.BR-H2k H2-T18a/SgSnJJrep and B10.BR-H2k H2-T18a/SgSnJ (drifted) mouse strains. Bone marrow cells from ten male mice of each strain, aged 10-12 weeks, were harvested. One pooled sample was analyzed for each strain.
Project description:We investigated the inflammatory gene profile of surgically-repaired lacerated muscles during the recovery phase and examined if it was influenced by the integrity/injury of the IM, intramuscular nerve using 3 lacerated rat muscle models: DN, where the IM-nerve was concomitantly cut; RN, where the IM-nerve was crushed but leaving an intact nerve sheath; and PN, a control where the IM-nerve was preserved intact. After 2, 4 and 8 weeks, the animals were sacrificed and their gastrocnemius muscles were removed and total RNA was extracted with Qiagen RNeasy Fibrous Tissue Mini Kit according to manufacturers instructions. Biotin-labelled cRNA probes were synthesized from total RNA by using a TrueLabeling-AMP Linear RNA amplification kit (SABiosciences Corp, Frederick, MD). The labeled cRNA probes were hybridized to oligonucleotide fragments spotted on the gene array membranes. Membranes were washed to remove any unincorporated probe and incubated with alkaline phosphatase-conjugated streptavidin (AP-streptavidin). Relative expression levels of specific genes were detected from signals generated by chemiluminescence from the alkaline phosphatase substrate, CDP-Star. The luminizing blots were used to expose X-ray films and quantified by spot densitometry with the aid of GEArray expression analysis suite (SABiosciences Corp, Frederick, MD). The abundance of each transcript was normalized to the normal un-operated muscle of the opposite limb, and to the housekeeping gene markers on each array (Aldoa, GAPdH and BAS2C). The results were presented as log2-fold expression with PN used as a control, to compare the two different types of nerve injuries (RN and DN).
Project description:The hepatocyte growth factor (HGF)/c-Met signaling pathway is known to mediate vascularization. We have previously demonstrated that expression of a human HGF transgene in the small airways produced mice (HGF TG) that were more susceptible to the tobacco carcinogen, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). We also have observed that HGF TG mice display significantly enhanced vascularization in the lungs that increases over time compared to wild-type (WT) littermates. To analyze which genes might contribute to increased vascularization from HGF overexpression in the airways, RNA and protein were isolated from whole lungs of individual HGF TG and WT adult mice. We profiled the mRNA expression of several hundred genes representative of six biological pathways involved in transformation, angiogenesis, and tumorigenesis using two commercial microarrays. Significant changes in expression over a 1.5-fold boundary were also observed in lung tumors derived from NNK-treated HGF TG mice. Lung tumors were induced by exposing mice to four weekly i.p. injections of 3mg NNK (15μg/μl) over 2 weeks. Whole lungs from control untreated animals were dissected after sacrifice at 10, 20, or 40 wks of age, and NNK induced lung tumors were dissected from the animals at 20 or 40 weeks of age. Total RNA was extracted from whole lung or isolated tumors from HGF TG or WT mice using TRIzol reagent and the Array Grade Total RNA Isolation Kit. The cDNA was generated and labeled using the TrueLabeling-AMP Linear RNA Amplication Kit. RNA was analyzed from 28 mice in total. The angiogenesis array was used to analyze 10 samples taken from 40 week old mice (HGF TG untreated [n=4], WT untreated [n=4], HGF TG NNK treated [n=1], WT NNK treated [n=1]) and 6 samples from 20 week old mice (HGF TG untreated [n=2], WT untreated [n=2], HGF TG NNK treated [n=1], WT NNK treated [n=1]). The cancer gene array was used to analyze 8 samples taken from 40 week old mice (HGF TG untreated [n=2], WT untreated [n=2], HGF TG NNK treated [n=2], WT NNK treated [n=2]) and 4 samples from 20 week old mice (HGF TG untreated [n=2], WT untreated [n=2]).
Project description:Hepatectomy generally offers the best chance of long-term survival for patients with hepatocellular carcinoma (HCC). Many studies have shown that hepatectomy accelerates tumor metastasis, but the mechanism remains unclear. In this study, palliative hepatectomy was performed in an orthotopic nude mice model of HCC (MHCC97H) with high metastatic potential. Using Human Tumor Metastasis Microarray, we screened the metastasis-related genes in tumor tissues following palliative resection, and found that Metastasis suppressor 1 (MTSS1) located in the central position of gene function net of residual HCC; MTSS1 was up-regulated in residual tumor after palliative resection. Further studies found that MTSS1 enhanced the metastasis of residual HCC. In hepatitis B-related HCC patients undergone palliative hepatectomy, those with higher MTSS1 mRNA expression accompanied by the activation of matrix metalloproteinase 2 (MMP2) in residual HCC, had earlier residual HCC detection after hepatectomy and poorer survival when compared to those with lower MTSS1 level. In different cell lines, the levels of MTSS1 mRNA increased in parallel with metastatic potential. MTSS1 down regulation via siRNA decreased MMP2 activity, reduced invasive potentials of HCC by 28.9% in vitro, and averted the deteriorated lung metastatic extent in vivo. In conclusion, the poor prognosis of hepatitis B-related HCC patients following palliative hepatectomy associates with elevated MTSS1 mRNA expression; therefore, MTSS1 may provide a new research field for HCC diagnosis and treatment. Eighteen nude mice bearing HCC xenografts were randomized into three groups 14 days after orthotopic implantation: palliative resection group (mice received partial HCC resection with preservation of 2 mm tumor pedicles), sham operation group (mice only undergone an exposure of liver but without resection), and blank control group (mice without further surgical intervention). All mice were sacrificed by cervical dislocation 14 days following palliative resection based on pre-experimental results. The first time surgically removed HCC tissues were named tumor tissues T1; the second time surgically removed HCC tissues from sham operation group were named tumor tissues T2; Tissues from blank control group were named tumor tissues T3; Tissues from palliative resection group were named tumor tissues T4 that was residual HCC. Randomly selected 5 tumor specimens from each group were used for the screen of genes related to metastasis by microarray techniques.
Project description:[original title] Microarray analysis of DNA damage repair gene expression profiles in cervical cancer cells radioresistant to 252Cf neutron and X-rays. The aim of the study was to obtain stable radioresistant sub-lines from the human cervical cancer cell line HeLa by prolonged exposure to 252Cf neutron and X-rays. Radioresistance mechanisms were investigated in the resulting cells using SuperArray Oligo GEArray® Human DNA Damage Signaling Pathway Microarray. HeLa cells were treated with fractionated 252Cf neutron and X-rays, with a cumulative dose of 75 Gy each, over 8 months, yielding the sub-lines HeLaNR and HeLaXR.Gene expression patterns of the radioresistant sub-lines were studied through microarray analysis
Project description:From the analysis of all genes included in the array a number of individual genes were identified as dys-regulated in HIV-1 infected patients compared to healthy controls. In particular, increased CXCL13 expression in all individual patient samples. Abstract from publication: HIV-1 infection is associated with B-cell abnormalities such as hypergammaglobulinemia, poor immunisation responses and loss of serological memory. To determine whether altered expression of chemokine receptors and their ligands may play a role in B-cell dysfunctions during HIV-1 infection, the expression of CXCR4, CXCR5 and CCR7 receptors and their respective ligands on CD19+ B-cells were examined in HIV-1 infected patients and controls. We report a decreased CXCR5 expression on B-cells from patients (p<0.05), a phenomenon associated with a low CD4 T-cell count (<350 cells/µl). Interestingly, an increased expression of CXCL13, the ligand for CXCR5, was found in peripheral B-cells from HIV-1 infected patients. Moreover upon B-cell activation in vitro, CXCL13 was secreted in culture. In addition, CXCL13 positive B-cells were also found in the lymph nodes of HIV-1 infected patients, but not in control tissue. B-cell migration towards CXCL13, CXCL12 and CCL21, ligands for CXCR5, CXCR4 and CCR7, was also evaluated. In patients with a low CD4 Tcell count, migration towards all ligands was increased. Our findings indicate that altered expression of the chemokine receptor-ligand pair, CXCR5/CXCL13 may participate in the establishment of B-cell dysfunctions during HIV-1 infection. Total RNA was extracted from purified peripheral B-cells from 4 controls and 4 patients.
Project description:Effects of the matricellular protein CCN1 on macrophage inflammatory gene expression was tested in a macrophage cell line I-13.35. CCN1 (CYR61) is a matricellular protein that is highly expressed at sites of inflammation and wound repair. In these contexts, CCN1 can modify the activities of specific cytokines, enabling TNF-alpha to be cytotoxic without blocking NFkB activity and enhancing the apoptotic activity of FasL and TRAIL. Here we show that CCN1 supports the adhesion of macrophages through integrin alphaMbeta2 and syndecan-4, activates NFkB-mediated transcription, and induces a pro-inflammatory genetic program characteristic of classically activated M1 macrophages that participates in Th1 responses. The effects of CCN1 include upregulation of cytokines (TNFa, IL-1a, IL-1b, IL-6, IL-12b), chemokines (MIP-1a, MCP-3, Gro1, Gro2, IP-10), regulators of oxidative stress and complement (iNOS, C3), and downregulation of specific receptors (TLR4, IL-10rb) and anti-inflammatory factors (TGF-b1). CCN1 regulates this genetic program through at least two distinct mechanisms: an immediate-early response resulting from direct activation of NFkB by CCN1, leading to the synthesis of cytokines including TNFa and IP-10; and a delayed response resulting from CCN1-induced TNFa, which acts as an autocrine/paracrine mediator to activate the expression of other cytokines including IL-1b and IL-6. These results identify CCN1 as a novel component of the extracellular matrix that activates pro-inflammatory genes in macrophages, implicating its role in regulating macrophage function during inflammation. Macrophage cell line I-13.35 (from Tlr4-defective C3H/HeJ) was treated with purified recombinant CCN1 protein for 6 hr. Total RNA was isolated, converted to cRNA probes, and hybridized to a oligonucleotide array representing 113 murine inflammation-related genes (from SA Biosciences at www.sabiosciences.com, Cat. Number OMM-011.) The array is a nylon membrane based DNA microarray on which 60-mer oligonucleotides probes were printed. Gene expression profile of resting cells incubated in serum-free medium (containing albumin, BSA) was used as a control for basal levels of gene expression.
Project description:To clarify the role of monocyte in the pathogenesis of SSc, we analyzed gene and protein expression in monocyte from SSc and healthy controls in order to identify the molecules which were associated with pathogenesis of SSc. After the screeining using gene and protein expression profiles, functional role of the candidate molecules were further analyzed to clarify the role in the pathogenesis of SSc.