Supplemental Data
Files in this Data Supplement:
- Supplemental video file 2 (.avi, 143.1 MB) - Supplemental video file 2 (TEM.avi): Transmission electron microscopy aligned tilt series obtained from the same 1-yr old human skin prepared for confocal microscopy (Supplemental video file 1). Here, the skin was labeled en bloc with ADAMTS10 monoclonal antibody 287-3E2 followed by 1-nm enhanced immuno-gold. The movie was created by imaging a 300 nm thick section of labeled skin at a magnification of 17,500x using a FEI Tecnai G2 transmission electron microscope operated at 200KV, and the micrographs were recorded automatically using a bottom mounted FEI Eagle 2K CCD camera. The series consists of a total of 91 micrographs taken at a tilt increment of 2��� from 30��� to -30 ��� and an increment of 1��� from -30��� to -60��� and from +30 to +60���. Images were aligned using FEI Inspect-3D software. The series demonstrates ADAMTS10 localization to microfibril bundles adjacent to, but not directly intersecting the lamina densa of the DEJ.
- Supplemental video file 1 (.avi, 34.0 MB) - Supplemental video file 1 (confocal.avi): 3-dimensional reconstruction of thick section of 1-yr old normal human skin analyzed by dual fluorescence confocal microscopy (fibrillin-1 is red, ADAMTS10 is green, the overlap is yellow). Fibrillin-1 localizes to microfibrils extending into the dermis from the basal lamina of the epithelium. ADAMTS10 also localizes to microfibrils but is most intensely localized in a narrow region of the papillary dermis adjacent to the skin basement membrane (see also supplemental video file 2 below).