Figure 4.

LSD1 tethering interferes with the maintenance of CENP-A levels at the HAC. (A–C) CENP-A IF analysis of interphase 1C7 cells expressing (A) tetR-EYFP-LSD1^WT, (B) a catalytically inactive mutant fusion, LSD1^K661A or (C) a seven-fold lower level of tetR-EYFP-LSD1^WT 5 days after targeting the respective construct to the HAC centromere. Displayed cells represent the median arbitrary fluorescence unit (AFU) of the quantifications shown to the right. Arrowheads depict the HAC. For (C) a ‘day 7' median cell is shown. Scale bars: 5 μm. (Right panels) CENP-A IF signal quantification at HACs targeted by the indicated fusion constructs at the given time points after washing out of Dox. AFU values were normalized to the median value of the ‘day 1' time point. The dotted line indicates the 25% signal mark for orientation. Solid vertical lines represent the median. For 1C7-LSD1^WT cells, the corresponding full time course (days 1, 3, 5, 7) quantifications are shown in Supplementary Figure S3B, and day 1 and day 3 values are reproduced in Figure 3E.