Fig. 5

Effects of TALE-demethylase expression on the density of pericentromeric proteins. U2OS cells expressing the TALE-demethylase (top) or the catalytically inactive mutant (bottom). DNA is visualized using Hoechst, the TALE (shown in green) is revealed with an anti-HA antibody and HP1α (A), INCENP (B) or Aurora B (C) (shown in red) is revealed with a specific antibody. White arrows indicate TALE foci in each cell. Boxplots showing the signal at the TALE-demethylase foci (blue) (HP1α: n = 607 nuclei in 5 experiments, INCENP: n = 282 nuclei in 2 experiments, Aurora B: n = 153 nuclei in 2 experiments) or at the point mutant (orange) (HP1α: n = 308 in 5 experiments, INCENP: n = 130 in 2 experiments, Aurora B: n = 118 in 2 experiments). Signal in the foci is normalized by the global signal in the nucleus for HP1α, INCENP and Aurora B. The box represents 50% of data points and the whiskers extend up to 1.5 times the interquartile range, the horizontal bar represents the median. The dashed line represents a signal normalized equal to one. p values were computed using a Wilcoxon–Mann–Whitney test (*p < 0.05/**p < 0.01/***p < 0.001/****p < 0.0001). A single focal plane is shown for each cell. Scale bar, 5 µm