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. 2022 Mar 16;12(3):210381. doi: 10.1098/rsob.210381

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© 2022 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 6. — CDK cross-talk to regulate Cdk10 activity. (a) Immunoblot analysis of thio-phosphorylation (upper panel) or MBP (lower panel) of 1 µg MBP-Cdk10 after incubation with 0.2 µM of the indicated kinase for 60 min. Asterisks indicate GST-CycB1/GST-Cdk1 and GST-CycA2/GST-Cdk2 autophosphorylation. (b) Immunoblot analysis of thio-phosphorylation of 1 µg MBP-Cdk10 and MBP-Cdk10T196A after incubation with 0.05 µM Cdk1/CycB1 or Cdk5/p35NCK. Below, Ponceau S staining of the membrane. (c) Results of the mass spectrometric determination of phosphorylation sites of MBP-Cdk10 after 20 min incubation without any additional kinase or after incubation with Cdk1/CycB1, Cdk5/p35 or Cdk10/CycQ. The asterisk indicates phosphosites that were identified only after longer incubation times of 40 or 120 min.