mouse dpp4 cd26 antibody  (R&D Systems)

Journal: Genes & Development

Article Title: Igf2 regulates early postnatal DPP4 + preadipocyte pool expansion

doi: 10.1101/gad.352710.125

Figure Lengend Snippet: DPP4 + preadipocytes express significantly high levels of Igf2 . ( A ) MTT assay of primary adult scWAT preadipocytes cultured with 1% adult mice mouse serum and supplemented with or without 10 nM IGF2 for 7 days ( n = 5). ( B ) Reclustering of prewean preadipocytes only, shown as a UMAP plot. ( C ) Violin plot of Igf2 expression between two clusters identified in B ( left ) and bar graph of mRNA expression levels in DPP4 sorted cells from isolated prewean and adult SVCs ( right ) ( n = 3). ( D ) Violin plot of Dpp4 expression between two clusters identified in B ( left ) and bar graph of mRNA expression levels in DPP4 sorted cells from isolated prewean and adult SVCs ( right ) ( n = 3). ( E ) Fluorescent in situ hybridization by RNAscope of Igf2 (gray) and immunohistochemistry of DPP4 (green), WGA (red), and DAPI (blue) were performed on prewean scWAT isolated with skin ( n = 4). Scale bars: left , 500 µm; right , 25 µm. The tile scan images were taken with a 40× objective. (*) P = 0.0222, (***) P = 0.0002, (****) P < 0.0001. Data are shown as mean ± SEM.

Article Snippet: Cells were incubated with a DPP4 antibody (1:10; R&D Systems AF954) for 30 min, followed by addition of 15 μL of protein G magnetic beads (Miltenyi Biotec) for 30 min. MS or LS columns (Miltenyi Biotec) were calibrated with blocking buffer and loaded with cells.

Techniques: MTT Assay, Cell Culture, Expressing, Isolation, In Situ Hybridization, RNAscope, Immunohistochemistry

Journal: Genes & Development

Article Title: Igf2 regulates early postnatal DPP4 + preadipocyte pool expansion

doi: 10.1101/gad.352710.125

Figure Lengend Snippet: Loss of Igf2 in DPP4 + cells induces preadipocyte differentiation. ( A ) Dot plot of Ki67 expression in prewean and adult Dpp4 + preadipocytes. ( B ) mRNA expression of Ki67 in MACS-sorted DPP4 + and DPP4 − prewean and adult cells ( n = 3). ( C ) MTT assay of DPP4 + primary preadipocytes isolated from prewean and adult mouse scWAT ( n = 2). ( D ) Dot plot of Ki67 in Dpp4 + prewean preadipocytes comparing Igf2 -positive and Igf2 -negative cells. Igf2 was knocked down (siIgf2) in prewean DPP4 + primary preadipocytes. ( E ) mRNA expression of adipogenic markers ( n = 3). ( F ) Bright-field images of Igf2 knockdown and control differentiated prewean preadipocytes ( n = 4). Scale bar, 150 μm. ( G ) mRNA expression of adipogenic markers in DPP4 − cells ( n = 3). Data are shown as mean ± SEM.

Article Snippet: Cells were incubated with a DPP4 antibody (1:10; R&D Systems AF954) for 30 min, followed by addition of 15 μL of protein G magnetic beads (Miltenyi Biotec) for 30 min. MS or LS columns (Miltenyi Biotec) were calibrated with blocking buffer and loaded with cells.

Techniques: Expressing, MTT Assay, Isolation, Knockdown, Control