Project description:miRNA profiles of adipocyte-derived microvesicles (ADMs) on the Day 2-4 and Day 8-10 were compared. ADMs were prepared from the 48h-conditioned medium of 3T3-L1 adipocytes (Day 2-4 and Day 8-10) followed by RNA isolation.
Project description:mRNA profiles of adipocyte-derived microvesicles (ADMs) and their donor 3T3-L1 adipoyctes (Day 11) were compared. ADMs included RNA without typical 28S and 18S ribosomal RNA.
Project description:Comparison of gene expression level of 3T3-L1, PMEF and ES cell derived adipocytes to eWAT samples. We used microarray to investigate the gene expressional differences between different cellular models of adipocyte differentiation vs. in vivo eWAT tissues
Project description:Growing evidence indicates that PPARγ agonists, such as rosiglitazone (RSG,), induce adipose mitochondrial biogenesis. Using microarrays, we systematically analyzed nucleus-encoded mitochondrial gene expression in two common murine adipocyte models, 3T3 L1 and C3H/10T1/2 adipocytes, and aimed to further establish the direct role of RSG, and capture the temporal changes in mitochondrial gene transcription during this process. Experiment Overall Design: Fully differentiated 3T3 L1 and C3H/10T1/2 adipocytes were treated with RSG, or DMSO vehicle for 1, 2, 4, 7, 24, and 48 hrs, and total RNA was extracted for microarray analysis.
Project description:The LIM-domain-only protein FHL2 is a modulator of signal transduction and has been shown to direct the differentiation of mesenchymal stem cells toward osteoblasts and myocytes phenotypes. We hypothesized that FHL2 may simultaneously interfere with the induction of the adipocyte lineage. Therefore, we investigated the role of FHL2 in adipocyte differentiation using pre-adipocytes isolated from mouse adipose tissue and the 3T3-L1 (pre)adipocyte cell line. Here we report that FHL2 is expressed in pre-adipocytes and for accurate adipocyte differentiation, this protein needs to be downregulated during the early stages of adipogenesis. More specifically, constitutive overexpression of FHL2 drastically inhibits adipocyte differentiation in 3T3-L1 cells, which was demonstrated by suppressed activation of the adipogenic gene expression program as shown by extensive RNAseq analyses, and diminished lipid accumulation. To identify the protein-protein interactions mediating this repressive activity of FHL2 on adipogenesis, we performed affinity-purification mass spectrometry (AP-MS). This analysis revealed the interaction of FHL2 with the Nuclear factor of activated T-cells 5 (NFAT5), an established inhibitor of adipocyte differentiation. NFAT5 knockdown rescued the inhibitory effect of FHL2 overexpression on 3T3-L1 differentiation, indicating that these proteins act cooperatively. In conclusion, we present a new regulatory function of FHL2 in early adipocyte differentiation and revealed that FHL2-mediated inhibition of pre-adipocyte differentiation is dependent on its interaction with NFAT5.
Project description: this study analyzed the transcriptomic changes of 3T3-L1 adipocytes during differentiation, which is important for better understand the molecular mechanisms of obesity. The purpose of the current study is to provide a comprehensive understanding at the transcriptome level of adipocyte differentiation. The transcriptomic profies in 3T3-L1 adipocytes from different differentiation stage were examined using RNAseq technique.
Project description:Cebpa is a critical transcription factor gene for adipocyte differentiation and adipose tissue development. However, mechanisms controlling Cebpa expression during adipogenic differentiation remain largely unknown. Here, we generated the high-resolution chromatin interaction maps of Cebpa in 3T3-L1 preadipocytes (3T3-L1) and 3T3-L1 adipocytes (3T3-L1-AD) using circularized chromosome conformation capture coupled with next-generation sequencing (4C-seq), and characterized differences in their chromatin interactomes and chromatin status of the interaction sites during adipogenic differentiation. We performed a 4C-seq experiment on inguinal white adipose tissue (iWAT) to evaluate whether chromatin interaction between Cebpa-L1-AD-En2 and Cebpa promoters in 3T3-L1 adipocytes also exists in mouse adipose tissue.