Journal: Platelets

Article Title: Lymphatic blood filling in CLEC-2-deficient mouse models

doi: 10.1080/09537104.2020.1734784

Figure Lengend Snippet: CLEC-2 deficiency during chronic, but not acute, dermal vascular remodeling can lead to localized lymphatic vessel blood filling. Pf4-Cre CLEC-2 deficient mice and littermate wildtype controls were subjected to cutaneous reverse passive Arthus reaction (rpA) and skin biopsies were collected 4 h after reaction initiation. In neither genotype could blood-filled lymphatic vessels be identified by gross or microscopic assessment. (Ai) Representative images of rpA-challenged skin from Pf4-Cre CLEC-2 deficient mice and littermate wildtype controls. (Aii) Representative images of rpA-challenged skin sections from Pf4-Cre CLEC-2 deficient mice and littermate wildtype controls stained for podoplanin (brown) and hematoxylin (blue). Lymphatic vessels indicated by yellow arrows. Scale bar = 100 μm. Wildtype mice rendered thrombocytopenic by treatment of 2 μg/g of anti-GpIba rat anti-mouse monoclonal antibody mixture (R300) 18 h before rpA initiation had red blood cells in lymphatic vessels upon microscopic inspection. (Bi) Representative images of non-inflamed and inflamed areas of rpA-challenged skin sections from thrombocytopenic wildtype mice stained for podoplanin (brown) and hematoxylin (blue) and red blood cells (yellow). Lymphatic vessels indicated by yellow arrows. Scale bar = 50 μm. n = 4. (Bii) Quantification of the presence of red blood cells in podoplanin positive lymphatic vessels in rpA-challenged skin sections from thrombocytopenic wildtype mice. Pf4-Cre CLEC-2 deficient mice and littermate wildtype controls received an intradermal injection of B16F10 melanoma cells into back skin. After 12 days of tumor growth, skin was harvested. (C) Representative images of melanoma-challenged skin from Pf4-Cre CLEC-2 deficient mice and littermate wildtype controls and quantification of blood-filled tumor-associated lymphatic vessels (yellow arrows) in Pf4-Cre CLEC-2 deficient mice. n = 10.

Article Snippet: Anti-CLEC-2 rat anti-mouse monoclonal antibody (17D9, used for flow cytometry) and rat anti-mouse control monoclonal antibody were from BioRad.

Techniques: Staining, Injection