(A) The sequences of the three individual CAF-1 subunits are shown. Each bar represents an individual identical peptide observed in the protein between the compared samples, plotted as the difference in deuteron uptake between the CAF-1 and CAF-1-H3/H4 samples (i.e., difference = CAF-1-H3/H4 – CAF-1 only). The differences in deuteron uptake at 60’ are colored according to the legend. The 'cooler' colors (green, blue, and purple) represent an increase in apparent protection for the peptide in CAF-1-H3/H4 compared to the CAF-1 sample, whereas the 'warmer' colors (orange, yellow, and red) represent decreased apparent protection. Peptide coverage was approximately 60%, 80%, and 80% for Cac1, Cac2 and Cac3, respectively. (B) Differences in HX at 60’ were mapped on PHYRE2 models of Cac2 and Cac3. The coloring scheme is the same as for A, but amino acids with no coverage are colored dark gray to distinguish these residues from those that have coverage but did not exchange significantly. (C) The top panel shows five fold serial dilution analysis of strain CFY53 (cac1) with the vector pCac1 introduced that was either empty, expressed wild type Cac1 or Cac1 with the indicated amino acid changes. The bottom shows five fold serial dilution analysis of strain CFY54 (cac2) with the vector pCac2 introduced that was either empty, expressed wild type Cac2 or Cac2 with the indicated amino acid changes.