Project description:Nitidine Chloride(NC) were found to enhance IL-10 production in LPS-stimulated Bone-marrow dendritic cells(BMDCs ) ,while at the same time inhibit pro-inflammatory cytokines production, such as TNF- α and IL-6. BMDCs were treated with NC or vehicle following LPS stimulation to find out the influence of NC on BMDCs that regulate cytokines expression. This study indicated that NC regulate numerous gene expression, thus influence IL-10 and pro-inflammatory cytokines production in LPS-treated BMDCs.
Project description:To obtain the gene expression profiles of triple-negative breast cancer MDA-MB-231 cells perturbed by NCD, we treated MDA-MB-231 with either DMSO or 5 μM NCD for 24 hours. By comparing the expression profiles of DMSO-treated cells with those treated with Nitidine Chloride, we can identify the genes affected by Nitidine Chloride.
Project description:We used microarrays to detail the global programme of gene expression underlying Polo-like kinase inhibition with BI 2536 compound in mouse bone marrow-derived dendritic cells (BMDCs) stimulated with Toll-like receptor agonists LPS and poly(I:C). CD11c+ BMDCs were treated for 1 hour with BI 2536 (1µM) or vehicle control (DMSO) prior to stimulation with LPS or poly(I:C) for 4 h. Total RNA was extracted and prepared for hybridization on Affymetrix microarrays.
Project description:This study profiles transcriptional responses of male wild-type mouse endothelial cells following inflammatory stimulation with LPS relative to vehicle controls. Three independent male endothelial cell isolations were used as biological replicates. Cells were treated for 8 h with vehicle/starvation media or 500 ng/mL LPS, then lysed for RNA isolation and RNA-seq. media or 500 ng/mL LPS, then lysed for RNA isolation and RNA-seq.
Project description:To explore how nitidine chloride (NC) regulates biological functions of fibroblast-like synovioctyes (FLSs) from rheumatoid arthritis (RA) patients, we evaluated the transcriptome of NC-treated RA FLSs compared with untreated control using RNA sequencing analysis.
Project description:genes regualted by LPS or LPS+cAMP stimulation in BMDCs We used microarrays to identify genes that up-regulated by LPS+cAMP compared with just LPS.
Project description:genes regualted by LPS or LPS+cAMP stimulation in BMDCs; We used microarrays to identify genes that up-regulated by LPS+cAMP compared with just LPS. Experiment Overall Design: BMDCs were stimilated with LPS (10 ng/ml) in the presence or absence of cAMP (100 microM) for 3h. Specifically up-regualted gene by cMP was identified.
Project description:Gene expression was compared between CD11c+ bone marrow derived dendritic cells (BMDC) was compared between cells conditioned in 20nM 1,25(OH)2D3 (Vitamin D) or vehicle. A second analysis compared gene expression in 1,25(OH)2D3 or vehicle CD11c+ BMDCs which were matured in presence of 0.1ug/ml LPS.
Project description:Purpose: The goals of this study are to compare the different functions of manganese (II) and LPS on the activation and maturation of antigen presenting cells-BMDCs. Methods: BMDCs were generated by culturing bone marrow cells with 20 ng/ml of IL-4 (Genscript) and 20 ng/ml of GM-CSF (Genscript) at 37°C and 5% CO2 for 7 days.Then BMDCs were treated with MnCl2 (200 μM) or LPS (100 ng/ml) for 20 h. Then mRNAs were purified, sequenced and analyzed. Results: Using an optimized data analysis workflow, we compared 20339 transcripts in BMDCs. Manganese (II) induced 1677 transcripts expression with a fold change ≥1.5 and LPS induced 1555 transcripts expression with a fold change ≥1.5. Conclusion: Manganese (II) activates the production of type I IFNs and the maturation of BMDCs. But there are also some differences between functions of manganese (II) and LPS on BMDCs.
Project description:Dendritic cells (DCs) are a special class of leukocytes able to activate both innate and adaptive immune responses. They interact with microbes through germline-encoded pattern-recognition receptors (PRRs), which recognize molecular patterns expressed by various microorganisms. Upon antigen binding, PRRs instruct DCs for the appropriate priming of natural killer cells, followed by specific T-cell responses. Once completed the effector phase, DCs reach the terminal differentiation stage and eventually die by apoptosis. We have observed that following lipopolysaccharide (LPS)-stimulation the initiation of the apoptotic pathway in DCs is due the activation of NFAT proteins. Indeed, LPS induces Src-family kinase and phospholipase C (PLC)γ2 activation, influx of extracellular Ca2+ and calcineurin-dependent nuclear NFAT translocation. The initiation of this pathway is independent of TLR4 engagement, and dependent exclusively on CD14. According with this observation CD14-deficient DCs do not die following LPS stimulation. Nevertheless, CD14-deficient DC death following LPS activation can be restored by co-stimulating DCs with LPS and thapsigargin. Thapsigargin empties the intracellular calcium stores by blocking calcium pumping into the sarcoplasmic and endoplasmic reticulum and thereby activates plasma membrane calcium channels. This, in turn, allows an influx of calcium into the cytosol and NFAT activation. To identify the NFAT controlled apoptosis genes in LPS activated DCs we have performed a kinetic microarray analysis (0, 48 and 60 h) in conditions allowing or inhibiting NFAT activation. Four genes have been selected: Nur77, Gadd45g, Ddit3/Gadd153/Chop-10 and Tia1. To identify apoptosis genes selectively modulated by NFAT, a comparative kinetic (time points 0, 48 and 60 h) microarray analysis was performed in the following conditions: 1) wild type bone marrow derived DCs (wtBMDCs) stimulated with LPS; 2) CD14-deficient BMDCs stimulated with LPS; 3) wtBMDCs stimulated with LPS in presence of thapsigargin; 4) CD14-deficient BMDCs stimulated with LPS in presence of thapsigargin.
