Abnormal patterning of ear lymphatic capillaries in adult Bmp9-KO mice. (A) Adult ear lymphatic capillaries were stained for LYVE-1 (green) and CD31 (red). Note that the LYVE-1 staining overlies weak CD31 staining in the lymphatic vessels. Bar represents 300 µm. (B) Quantification of the mean LYVE-1–positive area expressed as percentage of total area. The lymphatic vessel area in the inner layer of the ear of adult mice was measured using Image J software on images corresponding to 3 different fields of whole ear skin. These regions were kept constant for all samples. (C) Quantification of the mean LYVE-1–positive lymphatic vessel section expressed in micrometers. To quantify ear skin capillary mean lymphatic size, 5 horizontal lines were evenly laid on the images, and the diameters of lymphatic vessels that crossed these lines with an angle above 45° were measured using Image J according to Zhou et al.31 Values are mean ± SE; n = 8 for each genotype. ***P ≤ .001, significantly different from WT pups by unpaired Student t test.
