1. FRAP and FLIP analysis of GFP-ADAR2 and GFP-ADAR1C-Term.
FRAP and FLIP analysis of GFP-ADAR2 and GFP-ADAR1C-Term. (A) HeLa cells expressing either GFP-ADAR2 or GFP-ADAR1C-Term were bleached in a selected nucleolus for 0.5 seconds. Images were taken before bleaching and at the indicated time points after the end of the bleach pulse. (B) HeLa cells expressing either GFP-ADAR2 or GFP-ADAR1C-Term were repeatedly bleached in a selected nucleolus at intervals of 4.5 seconds. Images were taken before bleaching and at the indicated time points after the end of the first bleach pulse. The bleached nucleolus is indicated by an arrow in the pre-bleached panels. The dashed lines indicate the contour of the HeLa cell nucleus. Corresponding quantitative data of fluorescence-recovery kinetics are plotted on the right-hand side of each set of images. For FRAP experiments (A), the fluorescence intensity in the bleached region was measured and expressed as a relative fluorescence recovery (normalized fluorescence; see Materials and Methods). For FLIP experiments (B), the fluorescence intensity was measured in an adjacent nucleolus (marked by a white circle) and normalized as in FRAP experiments. Bar, 10 μm.
Joana M. P. Desterro et al. J Cell Sci 2003;116:
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