Journal: Frontiers in cell and developmental biology

Article Title: Leukemia Inhibitory Factor (LIF) Overexpression Increases the Angiogenic Potential of Bone Marrow Mesenchymal Stem/Stromal Cells.

doi: 10.3389/fcell.2020.00778

Figure Lengend Snippet: FIGURE 8 | Pro-angiogenic potential of MSC_LIF conditioned medium in vivo. Mixture of Matrigel with indicated conditioned medium (MSC or MSC_LIF) or phosphate buffered solution (NEG_CTL) was injected into a mouse ventral area (n = 3). After 7 days, Matrigel plugs were measured for hemoglobin (Hb) using Drabkin’s solution. (A) Representative images and (B) quantification of Hb. (C) Immunofluorescent staining of frozen Matrigel section with antibodies against endothelial marker CD31 (green). Nuclei are counterstained with DAPI (blue). The two-way ANOVA test and the Bonferroni post-test were used to analyze the differences among the groups. Values are expressed as mean ± SD of at least three independent experiments. **p < 0.01; ***p < 0.001. Scale bars = 50 µm.

Article Snippet: Frozen Matrigel sections were stained with rat anti-mouse CD31 antibody (R&D system, 0.125 μg/mL in BSA/normal serum solution) followed by incubation with Alexa 488-conjugated anti-rat antibody (Molecular Probes, 10 μg/mL in PBS) as described previously by Ribeiro et al. (2019).

Techniques: In Vivo, Injection, Staining, Marker

Journal: PLOS One

Article Title: Conditioned medium of engineering macrophages combined with soluble microneedles promote diabetic wound healing

doi: 10.1371/journal.pone.0316398

Figure Lengend Snippet: A) H&E and Masson staining of the collected wound skin tissue in different groups on day 12. Immunohistochemical staining was employed to show platelet endothelial cell adhesion molecule-1 (CD31). B) The thickness of re-epithelialization on day 12. C) Collagen deposition of wound on day 12. D) Statistical analysis of the CD31 + at the wounds on day 12. * p < 0.05, **p < 0.01, ****p < 0.0001; ns not significant, black scale bar = 500 μm, white scale bar = 50 μm.

Article Snippet: This technique was employed to visualize platelet endothelial cell adhesion molecule-1 (CD31) (brown-yellow, DAB color development kit, Beijing Zhongshan Jinqiao Biotechnology Co., LTD), which is the marker of endothelial cells.

Techniques: Staining, Immunohistochemical staining

Journal:

Article Title: VEGF-A stimulates lymphangiogenesis and hemangiogenesis in inflammatory neovascularization via macrophage recruitment

doi: 10.1172/JCI200420465

Figure Lengend Snippet: Concomitant induction of HA and lymphangiogenesis in inflammatory corneal neovascularization. (A–F) Seven days after central, intrastromal suture placement (A), a robust angiogenic response (A; blood vessel [BV]) in combination with an influx of inflammatory cells (B [H&E] and C) can be seen biomicroscopically (A) and by using CD31 (PECAM1) immunostaining (D) of corneal flat mounts (green). The CD45+ inflammatory cell infiltrate (C) consists mainly of GR-1+ neutrophils (red) and F4/80+ macrophages (green). In addition to the CD31+++LYVE-1– blood vessels (D and E; green), there is parallel outgrowth of CD31+LYVE-1+++ lymphatic vessels (LV; D–F; red). Blood vessels do not react with the lymphatic vascular–specific hyaluronic acid receptor LYVE-1 (F). Magnification, ×20 (A), ×200 (B and F), ×400 (C and E), and ×100 (D).

Article Snippet: Briefly, corneal flat mounts were rinsed in PBS, fixed in acetone, rinsed in PBS, blocked in 2% BSA, stained with FITC-conjugated CD31 (platelet–endothelial cell adhesion molecule 1 [PECAM-1]) antibody overnight (1:100 dilution; Santa Cruz Biotechnology, Santa Cruz, California, USA), washed, blocked, and stained with anti-LYVE-1 (1:500 dilution; LYVE-1 is a lymphatic endothelium–specific hyaluronic acid receptor; D. Jackson, Oxford University, Oxford, United Kingdom) ( 22 , 30 ), which was visualized using a indocarbocyanine-conjugated secondary antibody (1:100 dilution; Jackson ImmunoResearch Laboratories, Westgrove, Pennsylvania, USA).

Techniques: Immunostaining

Journal:

Article Title: VEGF-A stimulates lymphangiogenesis and hemangiogenesis in inflammatory neovascularization via macrophage recruitment

doi: 10.1172/JCI200420465

Figure Lengend Snippet: Neutralization of VEGF-A inhibits HA and lymphangiogenesis. (A–F) A molecular trap designed to bind VEGF-A (VEGF TrapR1R2) completely inhibits both HA and lymphangiogenesis within 1 week after injury. Whereas mice receiving an intraperitoneal injection of Fc protein at surgery (Fc control) display robust angiogenesis (A, slit-lamp picture; B, CD31 staining) and lymphangiogenesis (C, CD31 and LYVE-1 staining) 1 week later, mice treated with a single injection of VEGF TrapR1R2 do not show HA (D and E; blood vessels are green) or lymphangiogenesis (F; lymph vessels are red). Magnification, ×100 (C–F). (G) Morphometric analysis of the nearly complete inhibitory effect of VEGF Trap on both HA and lymphangiogenesis (P < 0.001). Magnification (A and B), ×20.

Article Snippet: Briefly, corneal flat mounts were rinsed in PBS, fixed in acetone, rinsed in PBS, blocked in 2% BSA, stained with FITC-conjugated CD31 (platelet–endothelial cell adhesion molecule 1 [PECAM-1]) antibody overnight (1:100 dilution; Santa Cruz Biotechnology, Santa Cruz, California, USA), washed, blocked, and stained with anti-LYVE-1 (1:500 dilution; LYVE-1 is a lymphatic endothelium–specific hyaluronic acid receptor; D. Jackson, Oxford University, Oxford, United Kingdom) ( 22 , 30 ), which was visualized using a indocarbocyanine-conjugated secondary antibody (1:100 dilution; Jackson ImmunoResearch Laboratories, Westgrove, Pennsylvania, USA).

Techniques: Neutralization, Injection, Staining

Journal:

Article Title: VEGF-A stimulates lymphangiogenesis and hemangiogenesis in inflammatory neovascularization via macrophage recruitment

doi: 10.1172/JCI200420465

Figure Lengend Snippet: Importance of VEGF-A isoforms for lymphangiogenesis. (A–E) Double immunostaining CD31/LYVE-1 (blood vessels, green; lymphatic vessels, red) of corneal flat mounts of wild-type mice (A), VEGF-A164/164 transgenic mice (B), and VEGF-A188/188 transgenic mice (C) demonstrates significantly reduced HA (D; P < 0.05) and lymphangiogenesis (E; P < 0.05). Magnification, ×100 (A–C).

Article Snippet: Briefly, corneal flat mounts were rinsed in PBS, fixed in acetone, rinsed in PBS, blocked in 2% BSA, stained with FITC-conjugated CD31 (platelet–endothelial cell adhesion molecule 1 [PECAM-1]) antibody overnight (1:100 dilution; Santa Cruz Biotechnology, Santa Cruz, California, USA), washed, blocked, and stained with anti-LYVE-1 (1:500 dilution; LYVE-1 is a lymphatic endothelium–specific hyaluronic acid receptor; D. Jackson, Oxford University, Oxford, United Kingdom) ( 22 , 30 ), which was visualized using a indocarbocyanine-conjugated secondary antibody (1:100 dilution; Jackson ImmunoResearch Laboratories, Westgrove, Pennsylvania, USA).

Techniques: Double Immunostaining, Transgenic Assay