Fig. 5.

Simvastatin and U0126 suppress lymphangiogenesis in vivo. (A) Representative images of immunofluorescent stains reveal a lower density of lymphatic vessels, which are LYVE-1/CD31 double positive (white arrows) in the dermis overlying the implanted Matrigel plug. The lymphatic vessel (LV) number per mm epidermal basement membrane (BM, the interface between epidermis and dermis) (B) and the percent of dermal area covered by lymphatic vessels (D) were strongly reduced in simvastatin (2 mg/kg) and U0126 (15 mg/kg) treated mice compared to DMSO vehicle treated mice. No change in LV size was observed (C). S1P + V + F = S1P(2 μM) + VEGF-A(500 ng/ml) + bFGF(500 ng/ml) in Matrigel (MG). *** p < 0.001 in unpaired Student t-tests (seven animals with two plugs per group). (E) Representative micrographs of immunofluorescent LYVE-1 stains labeling lymphatic vessels and quantitative image analyses of LYVE-1 stains (F) reveal that simvastatin treatment significantly inhibited corneal lymphangiogenesis induced by suture placement (eight mice per group). * p < 0.05 in Mann-Whitney test. LV = lymphatic vessel. Scale bars represent 100 μm. Bars show mean ± SEM.