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. 2019 Feb 7;73(3):562–573.e3. doi: 10.1016/j.molcel.2018.11.025

Figure 2.

Figure 2

Interactions between Sld3, Cdc45, and Rad53 Are Required for Rad53-Dependent Phosphorylation of Sld3

(A) Scale diagram of budding yeast Rad53, Cdc45, and Sld3. The Cdc45 DHH phosphoesterase homology domain is in green. The Cdc45-binding domain (CBD) of Sld3 is in blue. The 2D mutant refers to Sld3 S306D and T310D.

(B, E, and G) Western blots of the indicated strains, released from G1 arrest in alpha factor (time 0) into 200mM HU.

(C and F) Western blots of the indicated strains arrested in nocodazole (0) and treated with 10 μg/ml 4-NQO for the indicated times.

(D) The indicated Cdc45 peptides were immobilized on beads and used in pull-down assays with glutathione S-transferase (GST) or Rad53-FHA1-GST. A Coomassie stained gel of the indicated percentage of input and pull-down is shown.