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Blood, Vol. 103, Issue 9, 3552-3561, May 1, 2004
WASp deficiency in mice results in failure to form osteoclast sealing zones and defects in bone resorption
Blood Calle et al.
103: 3552
Supplemental materials for: Calle et al, Vol 103, Issue 6, 3552-3561
Files in this Data Supplement:
- Video 1. Interference reflection microscopy films of SV129 osteoclasts plated on glass viewing chambers (MOV, 4 MB)
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Normal or WASp-null osteoclasts were plated on glass coverslips and mounted onto viewing chambers in culture medium. Interference reflection micrographs taken 10 seconds apart were collected for 20 minutes using a Zeiss Standard 18 microscope (Zeiss, Welwyn Garden City, United Kingdom) with an incident light fluorescence attachment. Exciter and barrier filters were removed from the LP420 reflector and replaced with a narrow band pass filter to isolate the 546-nm line of the mercury arc source. Coverslips with attached cells were observed using a Zeiss 63|[times] Neofluar Antiflex (Zeiss) oil immersion objective, NA 1.25. Images were collected digitally using in-house software and edited using Jasc Animation 3.0 software (Digital Workshop, Banbury, UK). Micrographs were taken 10 seconds apart, and 20 frames per second are displayed in the film. Normal osteoclasts formed very dynamic, well-organized actin rings that assembled and disassembled continuously.
- Video 2. Interference reflection microscopy films of WASp-null osteoclasts plated on glass viewing chambers (MOV, 4 MB)
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Micrographs were taken 10 seconds apart, and 20 frames per second are displayed in the film. WASp-null osteoclasts assembled more static actin rings. At the periphery of these rings, very dynamic adhesion points underwent rapid cycles of assembly and disassembly, although they failed to arrange in the same structures as normal cells.
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