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. Author manuscript; available in PMC: 2020 May 13.
Published in final edited form as: Cell Rep. 2019 Jul 23;28(4):1090–1102.e3. doi: 10.1016/j.celrep.2019.06.068

Figure 4. Ddc1 and Dpb11 regulate the enrichment of Mec1 S1964 autophosphorylation.

Figure 4.

(A) Checkpoint activity after the conditional depletion of Ddc1, Dpb11 or Dna2. Cultures of Ddc1-AID, Dpb11-AID and Dna2-AID strains together with a WT control were grown and split into two. One half was treated with IAA for 3h prior to induction of a DSB to allow the degradation of the AID-tagged proteins (+IAA). Samples were collected and probed for Rad53 and (B) for Ddc2-HA. (C) Strains that contain Ddc2-HA together with either Dpb11-AID-myc or Ddc1-AID-myc, and a WT control strain were treated with galactose for 3h, then cultures were split into two, one to be treated with IAA for 3h. Following Ddc2-HA immunoprecipitation, samples were probed for Mec1, phospho-Mec1-S1964 and HA, for detection of Ddc2 (upper panel). Samples collected from the lysates (bottom panel) were blotted for HA (for detection of Ddc2), myc (for detection of Ddc1 or Dpb11) and Rad53.

(D) Quantification of Mec1-S1964 phosphorylation after the depletion of Ddc1 or Dpb11. *p=0.02. Error bars represent SEM.