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Nuclear Factor B p65 Phosphorylation in Orthodontic Tooth Movement

Department of Orthodontics, 1600 SW Archer Road, Campus Box 100444, University of Florida College of Dentistry, Gainesville, FL 32610, USA

View larger version (46K): [in this window] [in a new window]   Figure 1. No change in levels of p65*536 was detected in RAW 264.7 cells related to stimulation with RANK-L. (A) RAW 264.7 cells were stimulated with vehicle or recombinant RANK-L for 5 days. Cultured cells that had been treated with RANK-L were visually inspected for the presence of giant multinucleated cells. Whole-cell extracts were then subjected to SDS-PAGE, blotted on nitrocellulose, and probed with antibodies to p65 or p65*536. There was no difference detected in the level of p65*536 staining depending on treatment. (B) p65*536 was detected surrounding and within the nuclei of RANK-L-stimulated RAW 264.7 cells. This experiment was typical of 3 separate experiments. Bar equals 20 µm.

View larger version (134K): [in this window] [in a new window]   Figure 2. Orthodontic tooth movement does not alter levels of total p65 in alveolar bone from rats. Representative sections of alveolar bone and periodontium associated with control or orthodontically stimulated incisors from rats stained for the presence of total p65. These are typical of staining detected for all controls and all time-points in 3 separate trials from 6 animals. The root and periodontal ligament (PDL) are indicated. The bars equal 50 µm.

View larger version (163K): [in this window] [in a new window]   Figure 3. Orthodontic tooth movement triggered a large increase in p65*536 in alveolar bone from rats. Representative sections of the alveolar bone underlying incisors orthodontically stimulated for 0, 3, and 12 hrs and stained for the presence of p65*536 are shown. Note the almost complete absence of staining at 0 hrs. Increased staining was detected by 3 hrs, and intense staining was observed at 12 hrs. The arrows point to a section in the 12-hour panel (bottom left) that was magnified in the bottom right panel. At high magnification, giant cells (osteoclasts) were detected and were stained intensely. The bar equals 50 µm in the two top panels and the bottom left panel, and 10 µm in the bottom right panel.

View larger version (9K): [in this window] [in a new window]   Figure 4. Scraping RAW 264.7 osteoclast-like cells triggers increased levels of p65*536. RAW 264.7 osteoclast-like cells were scraped from their substrate by means of a cell scraper, then prepared immediately for SDS-PAGE (0), or were allowed to settle for 1, 3, 6, or 12 hrs, and prepared for SDS-PAGE. After the proteins were separated, they were blotted on nitrocellulose and probed with either the anti-p65*536 antibody or the anti-p65 antibody. p65*536 was barely detected at time 0, but was present at high levels at subsequent time-points. The levels of total p65 were similar at all time-points.

Journal of Dental Research, Vol. 86, No. 6, 556-559 (2007)
DOI: 10.1177/154405910708600613


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