Fig. 2

Effects of TALE-demethylase expression on H3K9 methylation and H3K4me2. 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 H3K9me3 (A), H3K9me2 (B) or H3K9me1 (C) or H3K4me2 (D) (shown in red) is revealed with a specific antibody. Right panels: boxplots showing the signal normalized at the TALE-demethylase foci (blue) (H3K9me3: n = 215 nuclei in 3 experiments, H3K9me2/H3K9me1: n = 128 nuclei in 2 experiments, H3K4me2: n = 66 nuclei) or at the point mutant (orange) (H3K9me3: n = 129 nuclei in 3 experiments, H3K9me2/H3K9me1: n = 123 nuclei in 2 experiments, H3K4me2: n = 41 nuclei). Signal in the foci is normalized by the global signal in the nucleus. 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, 10 µm