IGF2 increases mBMMNC homing and neovascularization in the mouse Matrigel plug assay. C57BL/6 mice were injected with 0.6 mL Matrigel with or without IGF2 (200 ng) (n = 7 per group). After 1 day, CD34+ mBMMNCs from EGFP transgenic mice (5 × 105/mouse) were infused into the tail vein. Six days after infusion, mice were killed, and Matrigel plugs were excised and serially sectioned. (A) Representative Matrigel plugs. (B) Quantification of neovasculature formation by measuring hemoglobin levels in the Matrigel. (C) Plug sections were stained for infiltrating endothelial cells using anti-CD31antibody (Olympus CX31 RBSF microscope; 10×/0.25 NA and 40×/0.65 NA plan objectives). (D) Plug sections were used to identify ECs with anti-VWF antibody (red) and mBMMNCs with green fluorescence (Olympus IX81-ZDC microscope, LUCPL FLN 40×/0.60 NA lens). (E) Quantitative assessment of GFP+ mBMMNCs per field for each Matrigel sample. Data are the mean plus or minus SE. **P < .01 versus each mouse group.
