Fig. 5: Cdc14 inhibits resection by counteracting Cdk-dependent phosphorylation of Dna2. | Nature Communications

Fig. 5: Cdc14 inhibits resection by counteracting Cdk-dependent phosphorylation of Dna2.

From: Cdc14 phosphatase counteracts Cdk-dependent Dna2 phosphorylation to inhibit resection during recombinational DNA repair

Fig. 5

a Southern blot analysis of resection and DNA repair in cdc14-1 Dna2-AID PMV cells expressing a wild-type version of Dna2. After overnight culture in YP-Raffinose at 25 °C, HO was induced with galactose and the cultures were transferred to 33 °C. Six hours after HO induction, auxin was added to inactivate the endogenous Dna2-AID. Samples were taken before and after the addition of auxin, and genomic DNA was extracted, digested with StyI, and analyzed by Southern blot using probes located at increasing distances from the HO cleavage site as depicted in Fig. 2a. An ACT1 probe was used as a loading control. The graphs on the right represent the quantification of the averaged band signals from two Southern blots after normalization against their respective ACT1 signal and uncut T0 sample. U/R: Uncut/Repair. b Southern blot analysis of resection and DNA repair in cdc14-1 Dna2-AID PMV cells expressing a phospho-deficient version of Dna2. Southern blot was performed under the same experimental conditions as in a. The graphs on the right represent the quantification of the averaged band signals from two Southern blots after normalization against their respective ACT1 signal and uncut T0 sample. U/R: Uncut/Repair. c Southern blot analysis of resection and DNA repair in cdc14-1 Dna2-AID PMV cells expressing a phospho-mimetic version of Dna2. Southern blot was performed under the same experimental conditions as in a. The graphs on the right represent the quantification of the averaged band signals from two Southern blots after normalization against their respective ACT1 signal and uncut T0 sample. U/R: Uncut/Repair.

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