2.8 In vivo plasma extravasation experiment
Plasma extravasation was performed in 8-12 weeks old C57Bl/6 mice as previously reported (Wick et al. , 2018). Briefly, 10 nmol in 10 µL of BK-(1-9), BK-(1-7) or BK-(1-5) was injected intradermally in the hind paw and sterile saline was injected in the same volume (10 µL) on the contralateral paw as control. Immediately afterwards, Evans’ blue (Merck) solution at 2% was injected intraorbitally for measurement of plasma extravasation. After 30 minutes of peptide administration, mice were killed by cervical dislocation and footpads were dissected. The footpads were dried at 60°C for 24 hours and the tissue was weighted. Evans’ blue was extracted by formamide for 72 hours under constant stirring and then the absorbance of this final solution was measured by a spectrophotometer at λ = 620nm. Results were expressed as µg of Evans’ Blue per mg of dried tissue.