Journal:

Article Title: Lymphangiogenesis and angiogenesis: concurrence and/or dependence? Studies in inbred mouse strains

doi: 10.1096/fj.09-134056

Figure Lengend Snippet: VEGF-A, FGF-2 and VEGF-C-induced corneal angiogenesis and lymphangiogenesis in C57BL/6 and BALB/c mice. A) Pellets containing VEGF-A (200 ng), FGF-2 (100 ng), VEGF-C (400 ng), or vehicle control (PBS) were implanted into corneas of C57BL/6 and BALB/c mice. Corneal angiogenesis and lymphangiogenesis were examined 6 d after implantation. B) Double staining of corneal flat mounts for angiogenic (CD31, green) and lymphangiogenic (LYVE-1, red) endothelium. Scale bar = 400 μm. C, D) Quantitative analysis of angiogenesis (C) and lymphangiogenesis (D) in PBS-, VEGF-A-, FGF-2-, or VEGF-C-implanted corneas on d 6 (n=5–11). E) Quantitation of the number of lymphatic tips in corneas of VEGF-A-, FGF-2-, VEGF-C-, or vehicle control-implanted eyes (n=3–13). *P < 0.05; **P < 0.01.

Article Snippet: Matrigel plug assay Mice were injected subcutaneously with 0.2 ml Matrigel (356230; BD Biosciences) containing PBS, 1 μg FGF-2 (3139-FB; R&D Systems), or 2 μg VEGF-C (2179-VC; R&D Systems).

Techniques: Double Staining, Quantitation Assay

Journal:

Article Title: Lymphangiogenesis and angiogenesis: concurrence and/or dependence? Studies in inbred mouse strains

doi: 10.1096/fj.09-134056

Figure Lengend Snippet: Corneal lymphangiogenesis and conjunctival lymphatic vessels. Corneal flatmounts with lymphangiogenesis were examined on d 6 after implantation and divided into two groups, with or without preexisting lymphatic vessels in the conjunctiva. A) Double staining of corneal flatmounts for angiogenesis (CD31, green) and lymphangiogenesis (LYVE-1, red) with or without lymphatic tube structures in VEGF-A-implanted BALB/c mice. Blood vessels (arrowheads) and lymphatic vessels (arrows) in the conjunctiva are indicated. B) Frequency of the corneas with LYVE-1+ lymphatic tube structures in the pellet-implanted site of C57BL/6 and BALB/c mice. C) Quantitative analysis of lymphangiogenesis (white) and angiogenesis (black) in VEGF-A-implanted BALB/c corneas on d 6 (n=4 and 7). D) Photographs and immunohistochemistry of CD31 and LYVE-1 with or without conjunctival removal. E, F) Corneal angiogenesis and lymphangiogenesis were examined 6 d after PBS or VEGF-A implantation with or without conjunctival removal. Arrows indicate lymphatic hyperplasia around the excised area. G, H) Quantitative analysis of lymphangiogenesis (G) and angiogenesis (H) in PBS- or VEGF-A-implanted corneas on d 6 with or without conjunctival removal (n=4–7). I, J) Correlation between lymphangiogenic area (d 6) and limbus lymphatic ratio in VEGF-A (I) or FGF-2 implantation (J) (n=6–11). *P < 0.05; **P < 0.01. Scale bars = 400 μm.

Article Snippet: Matrigel plug assay Mice were injected subcutaneously with 0.2 ml Matrigel (356230; BD Biosciences) containing PBS, 1 μg FGF-2 (3139-FB; R&D Systems), or 2 μg VEGF-C (2179-VC; R&D Systems).

Techniques: Double Staining, Immunohistochemistry

Journal:

Article Title: Lymphangiogenesis and angiogenesis: concurrence and/or dependence? Studies in inbred mouse strains

doi: 10.1096/fj.09-134056

Figure Lengend Snippet: GF-induced angiogenesis and lymphangiogenesis in Matrigel plug assay. A) Double staining for angiogenic (CD31, green) and lymphatic endothelium (LYVE-1, red) of PBS-, FGF-2-, or VEGF-C-containing Matrigel in C57BL/6 and BALB/c mice, 10 d after subcutaneous injection of Matrigel. B, C) Quantitative analysis of LYVE-1+ area (red, B) and CD31+ area (green, C) in Matrigel (n=4). D) Double staining for lymphatic endothelium (podoplanin, red) and macrophages (CD11b, green) of Matrigel in C57BL/6 and BALB/c mice (d 10). E, F) Quantitative analysis of podoplanin+ area (red, E) and CD11b+ area (green, F) in Matrigel (n=4). G) Double staining for lymphatic endothelium (LYVE-1, red) and proliferating marker (Ki67, green) of FGF-2-containing Matrigel in BALB/c mice (d 10). H) Quantitative analysis of number of LYVE1+ Ki67+ cells in Matrigel (n=4). *P < 0.05; **P < 0.01. Scale bars = 100 μm (A, D); 50 μm (G).

Article Snippet: Matrigel plug assay Mice were injected subcutaneously with 0.2 ml Matrigel (356230; BD Biosciences) containing PBS, 1 μg FGF-2 (3139-FB; R&D Systems), or 2 μg VEGF-C (2179-VC; R&D Systems).

Techniques: Matrigel Assay, Double Staining, Injection, Marker