Fig. 3: Basal inhibition of PP2A-B56 is observed in directSLiM^LIE1 cells.

A Schematic illustrating kinetochore phosphosites on the BUB complex and KNL1 that are regulated by PP2A-B56. B–C Effects of PP2A-B56 inhibition on the levels of BUB1-pT609, BUB1-pT461 and BUB1 (B, left) or KNL1-pMELT, KNL1-pRVSF and KNL1 (B, right) or BUBR1-pT620 and BUBR1 (C) at unattached kinetochores, in nocodazole-arrested HeLa-FRT cells with/without directSLiM^LIE1 expression and treated with vehicle or rapamycin for 20 min. Kinetochore intensities from 20–30 cells, 2–3 experiments. D Levels of BubR1-pT620 at unattached kinetochores in nocodazole-arrested directSLiM^LIE1 cells treated with rapamycin for indicated times. Kinetochore intensities from 30 cells, 3 experiments E Heat map of phosphorylation sites upregulated after PP2A-B56 inhibition in directSLiM^LIE1 cells. Each row shows the z-scored intensity of a phosphorylation site (n = 170). Source data are provided as a Source Data file. Data information: Kinetochore intensities in B–C are normalized to directSLiM^LIE1 vehicle condition. Violin plots show the distributions of kinetochore intensities between cells. For each violin plot, each dot represents an individual cell, the horizontal line represents the median and the vertical one the 95% CI of the median, which can be used for statistical comparison of different conditions (see Methods).