Figure EV2. Phosphoproteomics on cells expressing the B56 inhibitor.

1. Live‐cell imaging of cells expressing doxycycline‐inducible B56 inhibitor. Cell were released from a thymidine block and followed into mitosis. Doxycycline was added at time 0 h.
2. Quantification of mitotic duration of cells from (A) Each circle represents a single cell and median time in mitosis indicated by red line. A representative result from at least three independent experiments is shown. At least 20 cells were counted per condition in the experiment shown. NEBD; nuclear envelope breakdown.
3. Comparison of log₂ ratios of B56‐dependent dephosphorylation sites versus other phosphorylation sites on the same protein.
4. IceLogo representation of over‐ and underrepresented amino acid residues surrounding phosphorylation sites for the down‐regulated phosphorylation sites from experiments presented in Fig 2A and B.
5. Venn diagram showing overlap of PP2A‐B56‐regulated sites in G1/S and mitosis (M).
6. Schematic of in vitro peptide phosphorylation assay set‐up corresponding to IceLogo of PP2A‐B56‐regulated sites in Fig 2F.
7. Schematic of LxxIxE inhibition experiment in mitotic lysate corresponding to IceLogo of PP2A‐B56‐regulated sites in Fig 2G.
8. Mitotic cell lysates treated with thiophosphorylated wt or S62A GST‐Arrp19 for 5 min followed by Western blotting with the antibodies indicated.

Source data are available online for this figure.