Laser scanning confocal microscopy and immunofluorescent microscopy of skin from male hBERK1 and matched HbA-BERK mice. (A-D) A montage of overlapping fields of Z-stack images (each 2.5-μm-thick) of 80- to 100-μm-thick sections of skin in HbA-BERK (A), hBERK1 (B), and BERK (C-D), showing vascular, lymphatic, and nerve architecture. Image shows CD31⁺ blood vessels (pseudo-colored red), PGP 9.5 immunoreactive nerves (pseudo-colored blue), and LYVE1⁺ lymphatic vessels (pseudo-colored green). Bar represents 250 μm (A-C) and 50 μm (D). The rectangular inset in panel D shows an enlargement of sprouting of nerve fibers, also seen in the encircled area. ep indicates epidermis; lv, lymphatic vessels; and np, nerve plexus. Each image represents 3 reproducible and similar images. Image acquisition information: FluoView FV1000 Laser Scanning Confocal BX61 Microscope (Olympus); 20×/0.70 (A-C), 40×/1.35 (D) oil objective lenses; In-built image acquisition system; Adobe Photoshop. (E) Morphometric analysis of epidermal and dermal thickness performed on hematoxylin and eosin-stained skin sections (shown in supplemental Figure 3; original magnification ×200) by substituting 1 unit for 0.5 μm as per the calibration of the micrometer. Data are shown as mean ± SEM of 9 measurements of 3 separate sections from 3 different mice. □ represents HbA-BERK mice; , hBERK1 mice; and ■, BERK mice. *P < .01.