Project description:Analysis of Immediate Early Response 2 (Ier2)-inducible NIH 3T3 cells after Ier2 induction with RheoSwitch ligand RSL-1. Results provide insight into the function of Ier2 in NIH 3T3 mouse embryonal fibroblasts. Immediate early genes, including Ier2, are rapidly induced in quiescent cells by proliferation and migration-inducing stimuli. Microarray gene expression profiling was performed to identify differentially expressed genes following overexpression of Ier2 in NIH 3T3-Ier2 inducible cells after 24 hour induction of Ier2.
Project description:Expression profiling of HepG2 human liver carcinoma cells and NIH 3T3 mouse fibroblasts after arsite treatment for 24h. RNA-seq data comprise 4 groups: NIH 3T3 mouse fibroblasts control and arsite treatment, and HepG2 human liver carcinoma cells control and arsenite treatment. Jena Centre for Systems Biology of Ageing - JenAge (www.jenage.de)
Project description:Adipocytes play a critical role in metabolic homeostasis. Here we report that Zbtb9 acts as a positive regulator of PPARγ transcriptional activity in 3T3-L1 mature adipocytes, and PPARγ target gene expression was decreased when Zbtb9 deficiency was induced by shRNA-mediated knockdown (KD), establishing Zbtb9 as a newly identified PPARγ cofactor. Given the central role of PPARγ in adipogenesis, we also sought to investigate the role of Zbtb9 in adipogenesis in 3T3-L1 cells. Surprisingly, lipid droplet formation was significantly increased in Zbtb9 KD cells compared with the control, as was the expression of adipogenic genes including Pparg, Adipoq, Fabp4, and Cd36. The transcriptome of Zbtb9-KD fibroblasts, prior to the initiation of differentiation showed that the E2F targets pathway was significantly upregulated compared to the control cells. In addition, E2F activity was increased in Zbtb9-KD fibroblasts using luciferase reporter assay. Accordingly, RB phosphorylation was enhanced in Zbtb9-KD cells. Collectively, these results demonstrate that Zbtb9 inhibits adipogenesis as a negative regulator of Pparg expression via altered pRB-E2F1 signaling. Together, our findings revealed complex cell-state dependent roles of Zbtb9 in adipocytes, identifying a new molecule that may be important in the pathogenesis and treatment of obesity and T2D.
Project description:To screen for specific circadian outputs that may distinguish the pacemaker in the mammalian suprachiasmatic nucleus (SCN) from peripheral-type oscillators in which the canonical clockworks are similarly regulated in a circadian manner, the rhythmic behavior of the transcriptome in forskolin-stimulated NIH/3T3 fibroblasts was analyzed and compared to that found in the rat SCN in vivo and SCN2.2 cells in vitro. Similar to the scope of circadian gene expression in SCN2.2 cells and the rat SCN, NIH/3T3 fibroblasts exhibited circadian fluctuations in the expression of the core clock genes, Per2, Bmal1 (Mop3), and Cry1 and 323 functionally diverse transcripts (2.6%), many of which were involved in cell communication. Overlap in rhythmically-expressed transcripts among NIH/3T3 fibroblasts, SCN2.2 cells and the rat SCN was limited to these clock genes and four other genes that mediate fatty acid and lipid metabolism or function as nuclear factors. Compared to NIH/3T3 cells, circadian gene expression in SCN oscillators was more prevalent among cellular pathways mediating glucose metabolism and neurotransmission. Coupled with evidence for the rhythmic regulation of the inducible isoform of nitric oxide synthase, the enzyme responsible for the production of nitric oxide, in SCN2.2 cells and the rat SCN but not in fibroblasts, studies examining the effects of a NOS inhibitor on metabolic rhythms in co-cultures containing SCN2.2 cells and untreated NIH/3T3 cells suggest that this gaseous neurotransmitter may play a key role in SCN pacemaker function. Thus, this comparative analysis of circadian gene expression in SCN and non-SCN cells may have important implications in the selective identification of circadian signals involved in the coupling of SCN oscillators and the regulation of rhythmicity in downstream cells or tissues. Keywords: Circadian time course