Evaluating Lymphatic Permeability in Mice

Ear lymphatic permeability was assessed as previously described^{21 (link)}. 7–10 days after the last dose of Tamoxifen, mice were anesthetized with Avertin. We intradermally injected the ears of anesthetized Rap1a^fl/fl, Rap1b^fl/fl; Prox1Cre^ERT2 mice or Rap1a^fl/fl; Rap1b^fl/fl mice with 3 μl of either histamine (10 mM, Sigma-Aldrich) or a combination of histamine (10 mM) and AM (10 μM) in saline with a 10 ul Hamilton syringe fitted with a 30 gauge needle. Three minutes later, the ears were subjected to intradermal injection of 3 μl 0.5% Evans Blue dye (Sigma-Aldrich) in saline. Images of the ears were obtained at 0 min and 10 min after dye injection.