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(A) A thermodynamic cycle illustrates how FEP is used to calculate ΔΔGfold for eight FOXO1 FKH mutants, with the L183R mutant shown as an example. (B) Correlation of ΔΔGfold values calculated using FEP (ΔΔGFEP) with ΔTm from TSA (black) and DSC (red). Uncertainty estimates for ΔΔGFEP are smaller than the marker width.
