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. 2016 Sep 22;1(15):e84095. doi: 10.1172/jci.insight.84095

Figure 6. Lymphatic regeneration requires infiltration of macrophages.

Figure 6

(A) Representative images of hind limbs following DT administration in control (PBS liposomes) and macrophage-depleted (clodronate liposome) animals (n = 5/group). (B) Quantification of increase in foot diameter from baseline in control and macrophage-depleted DT treated animals over time (*P < 0.001). (C) Representative H&E-stained cross sections of the distal hind limb harvested 4 weeks after DT administration in control and macrophage-depleted animals. Fibroadipose tissue of the hypodermis is outlined with dotted black lines and marked with an asterisk (scale bar: 1 mm). (D) Quantification of fibroadipose deposition as a percentage of the total cross-sectional area of the distal hind limb (*P < 0.001). (E) Left: Representative NIR imaging of control (top) and macrophage-depleted (bottom) animals 4 weeks after DT administration (n = 5/group). Middle and right: Whole mount dual immunofluorescence localization of LYVE-1 (middle) and LYVE-1/CD11b (right), demonstrating lymphatic vessels in the distal hind limbs of control (top) but not macrophage-treated (bottom) animals. Note the diminutive and hypoplastic appearance of lymphatics in macrophage-depleted mice (scale bar: 200 μm). (F) Quantification of dermal lymphatic vessels in hind limb tissue sections in control and macrophage-depleted animals 4 weeks after DT treatment (*P < 0.001). (G) Quantification of F4/80+ cells in hind limb tissue sections in control and macrophage-depleted animals 4 weeks after DT treatment (*P < 0.001). (H) ELISA for VEGF-A in hind limb protein from control and depleted mice at 3 weeks (*P < 0.01). (I) ELISA for VEGF-C in hind limb protein from each group of mice (*P < 0.01). 2-tailed Student’s t test.