Fig. 7
CCAN protein localization at the centromere depends on catalytic activity of SENP6. Knockdown of SENP6 can be rescued by reintroduction of wild-type SENP6, but not by reintroduction of catalytic dead SENP6. a, b U2OS cells stably expressing inducible shRNA-resistant GFP-tagged wild type (WT) (a) or catalytic dead (CD) SENP6 (b) were established. Expression of these constructs was induced by doxycycline for 24 h prior to transduction with lentiviruses encoding SENP6 shRNAs. Medium was replaced 1 day post infection. The next day, cells were seeded on coverslips and grown overnight. Subsequently, cells were fixed and stained with Hoechst to visualize DNA and CENP-A antibody. Panels show representative pictures of mitotic cells. Scatter plots show quantifications of the average CENP-A foci intensities per cell for two independent replicates. The data were statically analysed by two-sided t-test. *p < 0.05; **p < 0.01; ***p < 0.001, ****p < 0.0001. For a replicate 1: n (ctrl shRNA) = 14 cells, n (SENP6 shRNA1) = 14 cells; n (SENP6 shRNA2) = 15 cells; replicate 2: n (ctrl shRNA) = 14 cells, n (SENP6 shRNA1) = 15 cells; n (SENP6 shRNA2) = 15 cells. For b replicate 1: n (ctrl shRNA) = 14 cells, n (SENP6 shRNA1) = 13 cells; n (SENP6 shRNA2) = 15 cells; replicate 2: n (ctrl shRNA) = 15 cells, n (SENP6 shRNA1) = 15 cells; n (SENP6 shRNA2) = 14 cells. Dashed lines indicate areas of DNA. Scale bars = 5 µm. Error bars represent standard deviation. Source data are provided as a Source Data file
