Supplemental Material for Eisenstatt et al., 2020

Figure S1. Endogenous Flag-Cse4 is not mislocalized in cdc7-7 strains. Chromatin lysate from wild type (YMB10043) and cdc7-7 (YMB10040) strains with endogenous Cse4 tagged with Flag was immunoprecipitated against anti-Flag agarose. Input and IP samples were prepared for paired-end ChIP-seq and reads were aligned to the S. cerevisiae S288C reference sequence. A-G. Genome browser of input and ChIP samples for Chromosomes I, II, III, V, X, XI, and XV from wild type (top) and cdc7-7 (bottom) strains with Flag-Cse4. The CEN regions are indicated.
Figure S2. Overexpressed Flag-Cse4 is mislocalized in cdc7-7 strains. Chromatin lysate from wild type (YMB10044) and cdc7-7 (YMB10041) strains overexpressing Cse4 tagged with Flag was immunoprecipitated against anti-Flag agarose. Input and IP samples were prepared for paired-end ChIP-seq and reads were aligned to the S. cerevisiae S288C reference sequence. A-E. Genome browser of input and ChIP samples for Chromosomes II, III, X, XI, and XV from wild type (top) and cdc7-7 (bottom) strains with Flag-Cse4. The CEN regions are indicated.
Figure S3. Endogenous Flag-Cse4 is not stabilized in a cdc7-7 strain. Western blot analysis of protein extracts from wild type (YMB10043) and cdc7-7 (YMB10040) strains with Flag-Cse4 expressed from its own promoter were grown to logarithmic phase of growth. Cells were treated with cycloheximide (CHX, 50 µg/ml) and aliquots were taken at the indicated timepoints. Protein extracts were analyzed using Western blot analysis and blots were probed with anti-FLAG (Cse4) and anti-Tub2. (loading control). The graph shows the quantification of the levels of FLAG-Cse4 remaining after treatment with CHX relative to Tub2 from three independent experiments. Error bars represent SEM.
File S1 contains Table S1, which describes the strains used in this study, and Table S2, which lists the plasmids used.

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