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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) <t>Transwell</t> assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).
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Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) Transwell assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).

Journal: Bioactive Materials

Article Title: Reconstructing the ischemic osteogenic microenvironment through hierarchical scaffolds orchestrating Mg 2+ signaling and neuropilin-1–mediated angiogenesis

doi: 10.1016/j.bioactmat.2026.02.031

Figure Lengend Snippet: Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) Transwell assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).

Article Snippet: For the Transwell assay, BMSCs were seeded in the upper chambers of Transwell inserts (8.0 μm pore size; Servicebio, China), while different culture conditions were applied in the lower chambers according to the experimental groups.

Techniques: Wound Healing Assay, Transwell Assay, Migration, Immunofluorescence, Staining, Labeling, Western Blot, Expressing