Figure 7. CD25⁺ cell depletion does not improve lymphedema, decrease fibrosis, or augment lymphatic function. A.

Representative photograph of control or CD25+ depleted mice 6 weeks after tail superficial and deep lymphatic excision. B. Representative cross sectional histology and quantification of subcutaneous tissue thickness (brackets) in control and CD25+ cell depleted animals. C. Tail volumes in control and CD25+ cell depleted animals over the course of the experiment. CD25+ cell depletion was begun 2 weeks after surgery (arrow). D. Representative (of triplicate experiments) western blots from tail tissues for Th1 (IFN-y, Tbet), and Th2 (Gata-3, IL4) markers in control and CD25+ cell depleted animals 6 weeks after tail superficial and deep lymphatic excision. Quantification of band density relative to controls (fold change) is shown to the right. E. Lymphoscintigraphy and sacral lymph node uptake in control and CD25+ cell depleted mice 6 weeks after surgery. Representative heat map is shown to the right (white arrow = injection site; red circle = sacral lymph nodes). F. Scar index analysis (below) and representative photomicrographs of polarized light microscopic views (above) in control and CD25+ cell depleted animals (n = 5–7 per group) 6 weeks after surgery. G. Representative photomicrographs of type I collagen immunohistochemistry (above) and calculation of type I collagen staining in the dermis (positive pixels/mm²; below) in control and CD25+ cell depleted animals 6 weeks after surgery. H. Calculation of type I:type III collagen staining ratio in tail tissue sections from control and CD25+ cell depleted mice 6 weeks after surgery. I. Representative (of triplicate experiments) western blot analysis of a-sma, E-cadherin, type III collagen, pSMAD, and TGF-B1 in protein lysates obtained from tail tissues of control and CD25+ cell depleted animals 6 weeks after surgery. Quantification of band density relative to controls (fold change) is shown to the right.