Fig. 8. BzATP causes translocation of PKCβI from the cytosol to membrane. (A) PKCβI-EGFP was distributed throughout the cytosol of an osteoclast-like cell under control conditions, as seen at 0 seconds. Addition of BzATP at 120 seconds caused prompt translocation of fluorescence from the cytosol to the periphery of the cell, with recovery at later times. The lower panels are reconstructed z-stack images of the cell shown above at 0 and 250 seconds, showing that PKCβI-EGFP translocated exclusively to the upper (basolateral) membrane. Blue horizontal lines indicate the location of the confocal images shown above. Data are representative of five independent experiments. (B) Membrane localization plotted for cell shown in A, revealing that translocation of PKCβI to the membrane was transient and recovered. BzATP was added at the time indicated by the vertical broken line. Mean duration at half peak of the response was 101±15 seconds (determined in 12 osteoclast-like cells). The bar graph compares the response duration for PKC and PKCβI isozymes. ^*P<0.0001, analyzed by Student's t-test.