rat-anti-mouse mac2 primary antibody  (Cedarlane)

Journal: Scientific Reports

Article Title: Deletion of endothelial leptin receptors in mice promotes diet-induced obesity

doi: 10.1038/s41598-023-35281-7

Figure Lengend Snippet: Visceral adipose tissue inflammation. Immune cell subsets were analyzed in visceral adipose tissue (VAT) homogenates of End.LepR-WT and End.LepR-KO mice fed high-fat diet (HFD) for 16 weeks using flow cytometry. Representative dot plots after analysis of MHC-II and Ly6C are shown in ( A ), of CD11c and MHC11 in ( C ), the results of the quantitative analysis of CD45 + Lin − CD11b + Ly6C high CXCR1 + MHCII − cells in ( B ) and of CD45 + Lin − CD11b − CD11c + MHCII − in ( D ), respectively. Data are expressed as % of total living cells in VAT and were compared using One-Way ANOVA, Sidak’s multiple comparisons test. *p < 0.05, **p < 0.01 and ***p < 0.005. ( E ) Representative images after immunohistochemical analysis of macrophages in VAT of End.LepR-WT and End.LepR-KO mice fed HFD for 16 weeks. Scale bars represent 100 μm. The results of the quantitative analysis of the Mac2-immunopositive area are shown in ( F ), the number of crown-like structures (CLS) or nucleated giant cells per 100 adipocytes are shown in ( G ) and ( H ), respectively. Data were analyzed using Student’s t test. *p < 0.05 and **p < 0.01.

Article Snippet: Sections were incubated overnight at 4 °C with primary rat monoclonal antibody against Mac2 (CL8942AP; Cedarlane Laboratories; dilution, 1:400 in antibody diluent (Dako)).

Techniques: Flow Cytometry, Immunohistochemical staining

Journal: Scientific Reports

Article Title: Deletion of endothelial leptin receptors in mice promotes diet-induced obesity

doi: 10.1038/s41598-023-35281-7

Figure Lengend Snippet: Brain inflammation and energetic profiles of primary brain and VAT endothelial cells. Summary of findings after analysis of glycolysis and mitochondrial respiration in living primary endothelial cells isolated from brain (A,B) or visceral adipose tissue (VAT; C , D ) of female End.LepR-WT (n = 4–5) and End.LepR-KO (n = 7) mice fed standard laboratory diet (SD) or high-fat diet (HFD) for 16 weeks. Analyses were performed using the Seahorse XF96e Extracellular Flux analyzer. Results were normalized to the number of cells and are expressed as -fold change vs. End.LepR-WT mice. Data were analyzed using One-Way ANOVA, Sidak’s multiple comparisons test. Ns, non-significant. Representative immunofluorescence images of cryo-embedded brain sections from End.LepR-WT (n = 7–8) and End.LepR-KO (n = 7) mice, fed high-fat diet for 16 weeks, after staining with antibodies against CD206 (green) and CD31 (red) ( E ) or against Mac2 (green) and VEGF (red) ( H ). Size bars represent 20 µm. Results after quantification of the area immunopositive for CD206 ( F ), CD31 ( G ), Mac2 ( I ) or VEGF ( J ), expressed per total area of 40 × microscopic fields. Data were analyzed using Student’s t test. *p < 0.05 and **p < 0.01.

Article Snippet: Sections were incubated overnight at 4 °C with primary rat monoclonal antibody against Mac2 (CL8942AP; Cedarlane Laboratories; dilution, 1:400 in antibody diluent (Dako)).

Techniques: Isolation, Immunofluorescence, Staining