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Table 1.

Translocation of PKCα to the basolateral membrane of osteoclasts

Treatment Extracellular Ca2+ % Cells exhibiting translocation Number of cells tested Number of independent transfections
Osteoclast-like cells derived from RAW 264.7      
   BzATP   +   88   77   14  
   PMA   +   100   36   8  
   BzATP   –   0   14   5  
   PMA   –   67   12   5  
Osteoclasts derived from bone marrow of wild-type mice      
   BzATP   +   85   13   3  
   PMA   +   100   3   3  
Osteoclasts derived from bone marrow of P2xr7–/– mice      
   BzATP   +   0   10   3  
   PMA   +   100   10   3  
Treatment Extracellular Ca2+ % Cells exhibiting translocation Number of cells tested Number of independent transfections
Osteoclast-like cells derived from RAW 264.7      
   BzATP   +   88   77   14  
   PMA   +   100   36   8  
   BzATP   –   0   14   5  
   PMA   –   67   12   5  
Osteoclasts derived from bone marrow of wild-type mice      
   BzATP   +   85   13   3  
   PMA   +   100   3   3  
Osteoclasts derived from bone marrow of P2xr7–/– mice      
   BzATP   +   0   10   3  
   PMA   +   100   10   3  

Membrane localization of PKCα-EGFP was quantified from fluorescence intensity profiles of confocal images as described in the Results. Osteoclasts were stimulated with BzATP (150 μM) or phorbol myristate acetate (PMA; 10 μM) and, after treatment for 0.5-3 minutes, images were analyzed. Standard bath Ca2+ concentration was 2.4 mM unless EGTA (4 mM) was added to chelate extracellular Ca2+ where indicated

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