Deep 3T3-L1 adipocyte proteome at 10 days post-differentiation
Ontology highlight
ABSTRACT: Here we conducted a comprehensive analysis of the proteome of the widely used 3T3-L1 murine adipocyte laboratory cell line at 10-days post-differentiation.
Project description:Differentiation of 3T3-L1 cells into adipocytes involves a highly orchestrated series of events including clonal expansion, growth arrest and terminal differentiation. The mechanisms coordinating these different steps are not yet fully understood. Here we investigated whether micro (mi)RNAs play a role in this process. Microarray analysis was performed to detect miRNA expression during 3T3-L1 preadipocyte differentiation. Several miRNAs, including let-7, were up-regulated during 3T3-L1 adipogenesis. Ectopic introduction of let-7 into 3T3-L1 cells inhibited clonal expansion as well as terminal differentiation. The mRNA encoding high mobility group AT-hook 2 (HMGA2), a transcription factor that regulates growth and proliferation in other contexts, was inversely correlated with let-7 levels during 3T3-L1 cell adipogenesis, and let-7 markedly reduced HMGA2 concentrations. Knockdown of HMGA2 inhibited 3T3-L1 differentiation. These results suggest that let-7 plays an important role in adipocyte differentiation and that it does so in part by targeting HMGA2, thereby regulating the transition from clonal expansion to terminal differentiation. 3T3-L1 cells were induced to differentiation into mature adipocytes using a canonical DMI cocktail. The time point at two days after confluency of 3T3-L1 was defined as day 0. Samples were collected at day 0, day 1, day 4, and day 7. The expression of microRNAs at day 1, day 4, and day 7 was compared to that of day 0.
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:Insulin is a potent regulator of protein metabolism. Here we describe a time-resolved map of insulin-regulated protein turnover in 3T3-L1 adipocytes using metabolic pulse-chase labelling and high-resolution mass spectrometry.
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:Trans-10, Cis-12 conjugated linoleic acid (t10c12 CLA) causes fat loss in mouse 3T3-L1 adipocyte tissue culture. The early transcriptome changes were analyzed using high-density microarrays to better characterize the signaling pathways responding to t10c12 CLA. Their gene expression responses between 4 to 24 hr after treatment showed a common set of early gene expression changes indicative of an integrated stress response (ISR). Experiment Overall Design: Mouse 3T3-L1 RNA for each time point was isolated from control and treatment samples for analysis on microarrays with two to four biological reps.
Project description:The aim of this study was to investigate the molecular basis underlying the response of 3T3-L1 adipocytes to forced expression of UCP1. Keywords: adipocyte, Uncoupling protein 1, microarray