A montage of overlapping fields of z-stack images (each 2.5-μm-thick)of 80- to 100-μm-thick sections of skin showing the expression of mediators of pain in skin. HbA-BERK (A), hBERK1 (C), and BERK (E). Image shows CD31⁺ blood vessels (pseudo-colored green), SP immunoreactivity (pseudo-colored red), and CGRP immunoreactivity (pseudo-colored blue). Magenta color in panels C and E is indicative of colocalization of CGRP (blue) and SP (red). The inset in panel E shows localized, dense immunoreactivity for SP in BERK mice. Bar represents 100 μm. ep indicates epidermis. (B,D) Coexpression of MOR and CD31⁺ on blood vessels in HbA-BERK (B) and hBERK1 (D) mice. Image shows CD31⁺ blood vessels (pseudo-colored red), MOR immunoreactivity (pseudo-colored green), and 4,6-diamidino-2-phenylindole⁺ nuclei (pseudo-colored blue). Yellow represents coexpression of MOR on blood vessels. Original magnification ×150; scale bar represents 250 μm. Arrows indicate MOR expression in the epidermis and vasculature. ep indicates epidermis. Each figure is representative of images from skins of 3 different mice. Image acquisition information: FluoView FV1000 Laser Scanning Confocal BX61 Microscope (Olympus), 20×/0.70 oil objective lens, In-built image acquisition system, Adobe Photoshop (panels A,C,E); Olympus IX70 microscope, 15×/0.45 objective lens, DP70 digital camera and DP70 Manager software (Olympus), Adobe Photoshop (panels B,D).