*2.14. E*ff*ects on Vascular Permeability*

Changes in vascular permeability were assessed by visualizing extravasation of Evans Blue dye into the extravascular compartment of the skin [2,24,25]. Briefly, groups of five mice (per condition) were treated with the different types of inhibitors (as previously described in Pharmacological Treatments item). A dilution of the dye in PBS solution (30 mg/kg) was injected intravenously (100 μL) prior to intradermal injection of LiRecTCTP (10 μg) or PBS (negative control) (50 μL). After 60 min, animals were euthanized (intraperitoneal injection of ketamine 30 mg/kg and xylazin 5 mg/kg), and dorsal skin was removed for visualization of dye leakage and photographed. The patches of skin were excised and incubated in 2 mL of formamide at room temperature for five days, after which the absorbance of the resulting supernatant was measured at 595 nm. Results are of one representative experiment from three independent biological replicates (data shown in Figure 4). Alternatively, groups of five mice (per condition) were injected intravenously (100 μL) with a dilution of Evans Blue in PBS solution (30 mg/kg) prior to intradermal injection of LiRecTCTP (5 μg and 10 μg), LiRecDT1 (1 μg), recombinant GFP (10 μg) (negative control), and the same volume of PBS (50 μL) (negative control). After 60 min, animals were euthanized (as described above), and dorsal skin was removed for visualization of dye leakage and photographed (data shown in Figure 5).
