Journal: Molecular Metabolism

Article Title: Aregs-IGFBP3-mediated SMC-like cells apoptosis impairs beige adipocytes formation in aged mice

doi: 10.1016/j.molmet.2025.102125

Figure Lengend Snippet: Aging induces an elevation in Aregs and a reduction in SMC-like cells among ASPCs following cold exposure . (A) UMAP plot showing the clustering distribution of 58,714 cells from young and aged mice sWAT. (B) Dot plot displaying the expression levels and proportions of selected marker genes across different cell types. The size of the dot represents the percentage of cells expressing the gene, while the color intensity reflects the average gene expression level. (C) Stacked bar chart showing the proportion of cell types in sWAT at different time points (TN, 3 days, and 14 days post-cold exposure). (D) Ucp1 and Cd142 mRNA expression levels analyzed by qPCR in sWAT (n = 11). (E) Immunoblots for CD142, UCP1, TOMM20 and GAPDH in sWAT (n = 3). (F) Quantification of immunoblots of CD142, UCP1 and TOMM20, protein levels are normalized to GAPDH. (G) Immunohistochemistry fluorescence for UCP1 (red) and CD142 (green) (scale bar = 50 μm or 10 μm). Data represent mean ± SD, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Next, adipocytes were subjected to an overnight incubation at 4 °C with CD142 primary antibody (1:100, R&D Systems, AF3178).

Techniques: Expressing, Marker, Gene Expression, Western Blot, Immunohistochemistry, Fluorescence

Journal: Molecular Metabolism

Article Title: Aregs-IGFBP3-mediated SMC-like cells apoptosis impairs beige adipocytes formation in aged mice

doi: 10.1016/j.molmet.2025.102125

Figure Lengend Snippet: Aregs and IGFBP3 inhibit beige adipocyte formation. (A–E) All five groups of ASPCs underwent the same browning differentiation protocol. (A) Ucp1 mRNA expression levels analyzed by qPCR. (B) Immunoblots for UCP1, TOMM20 and GAPDH. (C,D) Quantification of immunoblots of UCP1 and TOMM20, protein levels are normalized to GAPDH. (E) Immunofluorescence staining for CD142 and Mitotracker for mitochondria (scale bar = 20 μm). Data represent mean ± SD, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Next, adipocytes were subjected to an overnight incubation at 4 °C with CD142 primary antibody (1:100, R&D Systems, AF3178).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Molecular Metabolism

Article Title: Two-step regulation by matrix Gla protein in brown adipose cell differentiation

doi: 10.1016/j.molmet.2024.101870

Figure Lengend Snippet: Global MGP deficiency disrupts brown adipogenic maturation . ( A ) Expression of the mesenchymal stem cell markers Cd44 , Cdh2 , Vim and Gata2 (left panel) and the adipose progenitor cell markers Cd34 , Cd142 , Icam1 and Pdgfra (right panel), in wild-type (WT) and Mgp -knockout (KO) iBAT, as determined by qPCR (n = 7 mice per group). ( B ) FACS analysis of adipose progenitor cells from the brown adipose stromal vascular fractions isolated from WT and KO mice (n = 5 mice per group), using antibodies to ICAM1, CD142, DPP4 and PDGFRA. ( C ) PDGFRA + DPP4+ double-positive cells from the stromal vascular fractions were cultured in adipogenic conditions for 12 days. The cells were stained with Oil Red O or collected for RNA analysis. The Oil Red O was extracted and quantified at 492 nM. Expression of Adipoq , Pparg and Prdm16 was determined by qPCR (n = 3; representative of 3 replicate experiments). Data are shown as mean ± SEM; (A and C) unpaired two-tailed Student's t test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The following antibodies were used: Perilipin1 (CST, 1:500), CD31 (R&D systems, 1:200), CD142 (Sino Biological, 1:50), MGP (Abcam, 1:2000), ICAM1 (Proteintech, 1:200), incubated at 4 °C overnight.

Techniques: Expressing, Knock-Out, Isolation, Cell Culture, Staining, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Two-step regulation by matrix Gla protein in brown adipose cell differentiation

doi: 10.1016/j.molmet.2024.101870

Figure Lengend Snippet: Single cell RNA-seq data analysis of GEO dataset GSE207707 . ( A ) t-Stochastic neighboring embedding (tSNE) plot of brown adipose stromal/stem cells (ASCs) and vascular cells. EC, endothelial cells; SMC, smooth muscle cells. ( B ) Scatter plot of Mgp expression in brown adipose ASCs and vascular cells. ( C ) Violin plots of the marker genes of brown adipose progenitor cells and pre-adipocytes. ( D ) Single-cell trajectory of brown adipose ASCs and the expression of marker genes along the trajectory. ( E ) Brown adipose ASCs ordered by pseudotime along the single-cell trajectory and gene expression dynamics of Mgp and Cd142/F3 in the ASC differentiation course.

Article Snippet: The following antibodies were used: Perilipin1 (CST, 1:500), CD31 (R&D systems, 1:200), CD142 (Sino Biological, 1:50), MGP (Abcam, 1:2000), ICAM1 (Proteintech, 1:200), incubated at 4 °C overnight.

Techniques: RNA Sequencing Assay, Expressing, Marker