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. 2020 Apr 30;39(11):e104419. doi: 10.15252/embj.2020104419

Figure 7. Mitotic Greatwall, ENSA and PP1 regulation are independent of cyclin B.

Figure 7

  1. Quantification of Greatwall, ENSA/ARPP19 and PP1 phosphorylation based on phospho‐proteomic data.
  2. Confirmation of ENSA/ARPP19 S67 phosphorylation in mitotic extracts from Ctr‐ and DIA‐treated cells, before and after ENSA/ARPP19 siRNA depletion.
  3. Dynamics of mitotic substrate dephosphorylation triggered by Cdk1 inhibition in control and DIA‐treated B1dd/B2ko cells. The cells were enriched in mitosis by Thymidine release and proTAME/Apcin treatment and treated with 1 μM flavopiridol to trigger mitotic exit. Dephosphorylation was measured by probing the immunoblots with anti Cdk1 substrate (anti‐pSP) antibody.
  4. Three repeats of the experiment in (C) were quantified and corrected based on the corresponding GAPDH intensity values. The bar plots show the mean of three experiments, and the standard deviation is indicated by error bars.
  5. Dynamics of Greatwall, PP1 and Cdc27 dephosphorylation following Cdk1 inhibition in proTAME/Apcin‐treated mitotic cell extracts from Ctr‐ or DIA‐treated cells. Cell extracts were prepared at the indicated times following treatment with 1 μM flavopiridol and analysed by immunoblotting (* indicates non‐specific band).