Project description:Epidydimal WAT from adipose tissue Pgam1 deficient mice shows the phenotype of beiging. We used single cell RNA sequencing (scRNA-seq) to characterize the beiging cells in gWAT.
Project description:To analyze the gene expression profile of BAT and gWAT from Pgam1 depletion mice, we performed whole genome microarray expression profiling using brown adipose tissue (BAT) and gonadal white adipose tissue (gWAT) from adipose tissue-specific Pgam1 knockout (KO) mice.
Project description:Obesity has become a global health problem. Brown adipose tissue (BAT), specialized for energy expenditure through thermogenesis, potently counteracts obesity. Recently, BAT is also identified in human adults. We found that Lgr4 homozygous mutant (Lgr4m/m) mice display reduced adiposity and exhibit brown-like adipocytes in their WAT depots with higher expression of uncoupling protein 1 (Ucp1). Furthermore, Lgr4 ablation potentiates brown adipocyte differentiation from stromal vascular fraction (SVF) of epididymal WAT (eWAT) in vitro. We used microarrays to emxamine the gene expression profiles of the brown-like adipocytes differentiated from SVF of wild-type and Lgr4 mutant mice. We identified distinct gene expression profiles of these two groups. To demonstrate Lgr4 ablation can potentiate the differentiation of SVF from eWAT toward brown-like adipocytes in vitro, we isolated SVF from epididymal white adipose tissue (eWAT) of wild-type (WT) and Lgr4 mutant mice. We then plated SVF cells in 12-well plate, and differentiated them to brown adipocytes, followed by RNA extraction and hybridization with Affymetrix microarrays.
Project description:We found deletion of Foxp4 in SVF cells would inhibit beige adipocyte differentiation and thermogenesis.In order to investage the binding sites of Foxp4 on genome of SVF cells, we isolated SVF cells from ingunal adipose tissues and did Foxp4 and H3K27ac ChIP-Seqs after two days' browning differentiaion.
Project description:We used microRNA expression analysis to identify which microRNAs are expressed in the stromal vascular fraction (SVF) of visceral white adipose tissue (visWAT) to identify of microRNA expression patterns are changed upon CL-316,243 treatment Agilent mouse miRNA microarray (Cat. No. G4872A-046065, Agilent Technologies) was performed on the SVF of visWAT of 8-week-old C57/BL6 mice treated with CL 316243 or Vehicle (PBS) for three days. Isolated miRNA and total RNA was used for array analysis by Shanghai Biotechnology Corporation. Normalization and analysis for differentially expressed genes were performed using robust multi-array analysis and significance analysis of microarrays (SAM) via R statistical software packages “oligo” and “samr”. Several up- and downregulated microRNAs could be identified and were further varified with qPCR and compared with mRNA expression levels of their main targets.
Project description:We previously derived a doxycycline-inducible (Tet-On) lineage-tracing model that allows for the indelible labeling of Pdgfrb-expressing perivascular cells in adipose tissue of adult mice (PdgfrbrtTA; TRE-Cre; Rosa26RmT/mG; herein, “MuralChaser mice”). Prior to exposing animals to doxycyline, all cells within the stromal-vascular fraction (SVF) of adult gonadal WAT (gWAT) express membrane tdTomato from the Rosa26 locus. Following 9 days of exposure to doxycycline-containing chow diet, Cre-mediated excision of the loxP-flanked tdTomato cassette occurs in Pdgfrb-expressing cells, and membrane-bound GFP (mGFP) expression is constitutively activated. We set out to test the hypothesis that Pdgfrb-expressing perivascular cells in gonadal visceral WAT of adult mice are heterogeneous, with subpopulations harboring functionally distinct phenotypes. To this end, we performed single cell RNA-sequencing of mGFP+ cells isolated from gWAT of lean (chow fed) 8 weeks-old male MuralChaser mice following 9 days of doxycycline exposure.
Project description:RNA sequencing from isolated MMP3+ cells in subcutaneous WAT SVF showed higher expression of genes associated with beige progenitors.