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Figure 8


Fig. 8. Protein exchange at PML NBs is dependent on ATP. (A) FRAP experiments were performed on the same GFP–PML-I-expressing U-2 OS cells before (37°C) or after treatment with 10 mM sodium azide/50 mM 2-deoxyglucose for 1 hour (37°C/{Delta} ATP). Similar experiments were performed at ambient temperature, without or in the presence of ATP-depleting drugs (22°C or 22°C/{Delta} ATP, respectively). Images represent maximum intensity projections from 3D time-lapse FRAP data sets. Arrows indicate bleached regions that contain a PML NB; these areas are also shown as enlarged insets. Scale bar, 5 µm. (B) FRAP experiments as described in A were performed on cells after the indicated times of ATP depletion. Fluorescence recovery after 10 minutes of the bleach pulse was quantified from 30 PML NBs (3 NBs x10 cells each) and displayed as mean ± s.d. (C) FRAP experiments as described in A were performed on cells that express the indicated GFP fusion protein after 60 minutes of ATP depletion. Fluorescence recovery at NBs after 10 minutes of the bleach pulse was quantified from ten cells each and displayed as the mean ± s.d.





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