Gene expression alteration in chondrocyte by HES1 overexpression
ABSTRACT: To investigate target molecules of HES1, we transduced GFP or HES1 into mouse chondrocyte cell line ATDC5, and performed the microarray analysis. HES1 overexpression increased inflammation-related genes including Il6 and Il1rl1. We established stable ATDC5 cells overexpressing GFP or HES1 by lentiviral transduction. We harvested mRNA from these cells, and performed microarray analysis.
Proceedings of the National Academy of Sciences of the United States of America 20150302 10
Notch signaling modulates skeletal formation and pathogenesis of osteoarthritis (OA) through induction of catabolic factors. Here we examined roles of Hes1, a transcription factor and important target of Notch signaling, in these processes. SRY-box containing gene 9 (Sox9)-Cre mice were mated with Hes1(fl/fl) mice to generate tissue-specific deletion of Hes1 from chondroprogenitor cells; this deletion caused no obvious abnormality in the perinatal period. Notably, OA development was suppressed w ...[more]
Project description:To investigate loss-of-function of the C/EBP family members, we used A-CEBP which exerts a dominant-negative effect against all CEBPs. DOX-inducible overexpression of A-CEBP into mouse chondrocyte cell line ATDC5 increased expressions of early differentiation markers, decreased those of late differentiation markers. In addition, A-CEBP altered many genes related with skeletal development, cartilage, cell cycle, inflammation and apoptosis. We established stable ATDC5 cells which express GFP or A-CEBP by DOX induction. We started differentiation of these cells by ITS supplement immediately after DOX induction, harvested mRNA after 3 weeks, and performed microarray analysis.
Project description:Synovial fibroblasts contribute to the inflammatory temporomandibular joint under pathogenic stimuli. Synovial fibroblasts and T cells participate in the perpetuation of joint inflammation in a mutual activation feedback, via secretion of cytokines and chemokines that stimulate each other. IL-17 is an inflammatory cytokine produced primarily by Th17 cells that plays critical roles in the pathogenesis of numerous autoimmune and inflammatory diseases. Here, we investigated the roles of IL-17A in temporomandibular joint disorders (TMD) by using genome-wide analysis of synovial fibroblasts isolated from patients with TMD. We analyzed the gene expression profiles of synovial fibroblasts that were treated with or without IL-17A. IL-17 induced gene expression in synovial fibroblasts from human temporomandibular joint was measured at 4 hours after treated with IL-17A (10 ng/ml) and untreated control samples. This experiment used one donor sample.
Project description:To know whether the Nup98-HoxA9 affects global gene expression, we performed DNA microarray analysis using the following four clones: two independent Nup98-HoxA9 ES clones (clone#1 and clone#9), parental ES cells, and HoxA9-Ct ES clone. Two independent Nup98-HoxA9 ES clones (clone#1 and clone#9), and HoxA9-Ct ES clone, was compared with parental ES cells (reference sample).
Project description:To confirm the changed gene expression profiles in GPR15 knock out (KO) macrophages, we applied a SurePrint G3 Mouse Gene Expression service from Takara Bio Inc (Kusatsu, Shiga, Japan) to analyze gene expression profiles in lipopolysaccharide (LPS)-stimulated GPR15KO macrophages, comparing with wild type (WT) macrophages. Most significantly up-regulated genes in GPR15KO macropahges included Il6, Il17a, Il23, Tnfsf8, Il1b, Ifna2 and Ccnd2. On the contrary, several inflammation-related genes, including Ccl17, Itgax, Nrp1 and Rasgrf2, were down-regulated in WT macrophages, compared to GPR15 KO macrophages. Abdominal macrophages from WT and GPR15 KO mice were stimulated with PBS or LPS (100 ng/ml) for 4 hrs. Total RNA were extracted using a TRIzol-chloroform based method.
Project description:Hydrostatic pressure is one of the main mechanical stimuli cartilage cells are submitted to during joint loading. If moderate hydrostatic pressure is known to be beneficial to cartilage differentiation, excessive pressure, on the other hand, induces changes in cartilage similar to those observed in osteoarthritic cartilage. Therefore, the purpose of the experiment is to identify new target genes of high hydrostatic pressure in chondrocyte precursor cells.
Project description:To obtain comprehensive profile of gene expression following a pancreatectomy, we performed microarray analysis of genes expressed in islets of C57BL/6J mice 3 days after 60% pancreatectomy or sham operation. gene expression in mouse pancreatic islets was measured at 3 days after 60% pancreatectomy or sham operation.
Project description:To study the gene expression profiles of brown (BAT) and white (WAT) adipose tissues in wild type and LR11-deficeint mice. The four RNA sources, WT scWAT, Lr11 -/- scWAT, WT BAT and Lr11 -/- BAT, were prepared from subcutaneous WAT and BAT from wild-type mice and Lr11 -/- mice, respectively (n=3 each).
Project description:The human genome contains several homologous Small Leucine-Rich Protein (SLRP)-encoding genes, many of which orchestrate collagen fibril assembly. All SLRP gene products have not been identified or characterized. In this report, we describe a novel class IV SLRP - Chondroadherin-like (CHADL). Microarray analysis of CHADL knockdown in ATDC5 cells suggests collagen receptor-related changes, a pro-proliferative and pro-differentiation profile. Two samples in duplicates are analyzed. The two control samples are transfected with scrambled negative control shRNA.The two knockdown samples are transfected with shRNA specific for CHADL.
Project description:The initiation and/or progression of ossification of the posterior longitudinal ligament (OPLL) is associated with cyclic tensile strain, but the pathomechanism of OPLL remains unclear. Indian hedgehog (Ihh) and its related signaling are key factors in normal enchondral ossification. However, the relation of OPLL to Ihh is unclear. The purpose of this study is to investigate the contribution of mechanical strain to OPLL and the relation of Ihh to OPLL. Cultured posterior longitudinal ligament cells were subjected to 24 hours of cyclic tensile strain and then analyzed by microarray.