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Figure 6


Fig. 6. Close temporal relationship between the BzATP-induced rise of [Ca2+]i and PKC{alpha} translocation. (A) Simultaneous imaging of [Ca2+]i using the fluorescent Ca2+ indicator dye Fura-red (red) and PKC{alpha}-EGFP (green). Fura-red intensity (F) decreases with the rise of [Ca2+]i, so data are expressed as the ratio of initial intensity (F0) to F for a region of interest in the cytosol. BzATP was added at 50 seconds (vertical broken line) and PKC translocation follows the rise in [Ca2+]i. Data are representative of seven out of ten cells tested. (B,C) BzATP induced a repetitive translocation of PKC{alpha}-EGFP in some osteoclast-like cells. An osteoclast-like cell was stimulated with BzATP (150 µM, applied at 25 seconds) as indicated, inducing repetitive translocation of PKC{alpha} between the cytosol and membrane. A confocal slice through the cell is shown in B, with membrane localization plotted in C for the same cell. Responses are representative of repetitive PKC translocation seen in 18 out of 68 osteoclast-like cells that responded to BzATP.





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