Fig. 3. Treatment with the DNA demethylating agent 5-Aza-2-deoxycytidine does not significantly alter MECP2e2 dynamics. (A) Balb/c 3T3 cells were treated with 1 µM 5-Aza-2-deoxycytidine (5-Aza-dC) for 5 days then transiently transfected with the human MECP2e2-GFP construct driven by the CMV promoter. 48 hours post-transfection, cells were photobleached. The recovery profiles of 5-Aza-dC-treated and untreated cells in heterochromatic regions are shown. The slight leftwards shift in the recovery curve was noted. (B) Recovery kinetics of 5-aza-dC treated and vehicle (DMSO) control in euchromatic regions are shown and indicate no changes in mobility associated with drug treatment. (C) Although there was a slight shift in the recovery curve in heterochromatin, the differences in the t⁵⁰ values following 5-Aza-dC treatment were not statistically significant in either heterochromatin and euchromatin. The t⁵⁰ values of vehicle and 5-Aza-dC treated cells in heterochromatin are 20.9±10.2 and 13.3±6.1, respectively (P=0.0798). In euchromatin, the t⁵⁰ values of vehicle and 5-Aza-dC-treated cells are 0.13±0.01 and 0.13±0.008, respectively (P=0.7561). The number of nuclei photobleached in each group is shown within the bar.