Immunohistological Profiling of Skin Samples

Immunohistology was performed as described previously (Quan et al., 2010 (link)). Briefly, skin samples embedded in OCT were sectioned (7 µm), fixed in 2% paraformaldehyde, permeabilized with 0.5% Triton X-100 in phosphate-buffered saline (PBS), blocked with corresponding serum (5% in PBS), and incubated for one hour at room temperature with SDF-1 (R&D Research, Minneapolis, MN, USA), HSP47, langerin, CD31, CXCR4, and α-smooth muscle actin primary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA), followed by incubation with corresponding secondary antibodies for one hour at room temperature. Between steps, the slides were rinsed for 10 min in Tris buffered saline with 0.1% Triton-X-100 (TBST). All sections were lightly counterstained with haematoxylin. The slides were examined using a digital imaging microscope (Zeiss, Germany). Specificity of staining was determined by substituting corresponding isotype-control immunoglobulins for the primary antibodies.