Project description:The impaired aerobic glycolysis of monocyte/macrophage are the main features in sepsis that encountered immunosuppression. NLRC3, an inhibitory sensor, can diminish the Toll-like receptor 4 pathways of macrophages which is associated with sepsis-induced immunosuppression, but whether NLRC3 plays key role in regulating the immunosuppression of macrophage via interfering glycolysis still unclear. The aim of this study was to characterize the transcriptome alterations by which NLRC3 loss, and further investigate the role of the glycolysis in the macrophage with the development of sepsis. Bone marrow-derived macrophages (BMDMs) was isolated from the femurs and tibias of the NLRC3 WT (LysM-Cre-NLRC3fl/fl) and NLRC3ΔMac (LysM-Cre+ NLRC3fl/fl) mice and then was treated with or without LPS for 12 hours, and the macrophages mRNA profiles of five-weeks-old NLRC3 WT mice (WT macrophages) and of five-weeks-old NLRC3ΔMac mice (NLRC3-/- macrophages) were generated by deep sequencing, in triplicate, using Illumina 6000.
Project description:To investigate the plasticity of Lipolysaccharide (LPS) tolerance, we employed microarray profiling to analyse the gene expression profile in macrophage. Four macrophage populations were induced; Untreated macrophages (Control group), Acute response to LPS (LPS activation group), LPS tolerance (T – Tolerant group) and recovered (R = recovered macrophage group) Using transcriptional analysis we demonstrate that recovery from LPS tolerance (R – Recovery), as defined by cytokine gene expression, is associated with a global change in the transcriptional profile of macrophage. This data confirms that LPS tolerance is a transient state which results in induction of novel hybrid macrophage activation state with a unique transcriptional signature. Bone marrow derived macrophages were polarised into three activation states; Acute response to LPS (A), LPS tolerant (T) and recovered (R). Gene expression was measured at 4 hours post stimulation with LPS. Three independent experiments were performed to measure gene expression changes between each macrophage group.
Project description:To investigate the plasticity of Lipolysaccharide (LPS) tolerance, we employed microarray profiling to analyse the gene expression profile in macrophage. Four macrophage populations were induced; Untreated macrophages (Control group), Acute response to LPS (LPS activation group), LPS tolerance (T – Tolerant group) and recovered (R = recovered macrophage group) Using transcriptional analysis we demonstrate that recovery from LPS tolerance (R – Recovery), as defined by cytokine gene expression, is associated with a global change in the transcriptional profile of macrophage. This data confirms that LPS tolerance is a transient state which results in induction of novel hybrid macrophage activation state with a unique transcriptional signature.
Project description:In order to fully characterize emodin's effects on macrophage activation, peritoneal macrophages were stimulated with LPS+IFNg with or without emodin and gene expression was analyzed using a whole genome microarray. Emodin significantly attenuated the IFNg/LPS induced changes in a large percentage of responsive genes (31%) through inhibiting multiple signaling pathways. RT-qPCR was used to confirm the results in several genes associated with M1 macrophage activation including: TNF, IL6, IL1b, iNOS, MMP2, and MMP9. Three-condition, one-color experiment: Vehicle control, LPS-IFNg or LPS-IFNg-Emodin treated periferal WBC PMN samples: 4 biological replicates each.
Project description:By microarray gene expression profiling, we investigated whether glutathione (GSH) has a signaling role in the response of RAW 264.7 cells to lipopolysaccharide (LPS). GSH was depleted by prior incubation with 120 µM buthionine sulfoximine (BSO) for 24 h, then cells were treated with 10 ng/ml LPS for 2 or 6 h. DNA microarray analysis was performed in triplicate samples in control and GSH-depleted cells, in the presence or absence of LPS.
Project description:To investigate the effect of macrophages on mammary basal cells, we established a macrophage depletion mouse model by clodronate liposome (CL) intraperitoneal injection, and the mammary basal cells were collected by FACS sorting. We then performed RNA-seq analysis on clodronate liposome (CL) treated mammary basal cells compared to control.