Extended Data Fig. 7: DMS assay validations in 5-FC and cytosine media.

a) Location on the cytosine/5-FC landscape of the Fcy1 variants (shown as grey dots) selected for validations superimposed on the density plot presented in Fig. 2d. The circles used to define the three clusters are also shown: green for silent-like mutants, magenta for nonsense-like mutants, and blue for front minimum mutants. Mutants falling outside these clusters were classified as ‘other’ and encompass most outliers from the DMS screen. Variants with both high 5-FC and cytosine DMSscore are shown as squares: these outliers potentially escape the resistance-function trade-off. b) Spearman’s correlation between growth curve replicates in 5-FC media, n = 73 variants. c) Spearman’s correlation between growth curve replicates in cytosine media, n = 72 variants. Data collected from the cytosine media from the outlier (T86M) was excluded from downstream analysis. d) Linear least-squares regression fit and Spearman’s correlation between 5-FC growth rates measured in the 1st and 2nd rounds of validations for 17 mutants present in both growth curve assays. e) Linear least-squares regression fit and Spearman’s correlation between cytosine growth rates measured in the 1st and 2nd rounds of validations for 17 mutants present in both assays. f) Growth rate values of the 2nd round of validations scaled to the growth rate of the 1st round for the 17 mutants present in both assays. g) Spearman’s correlation between validation growth curves in 5-FC media and DMS 5-FC score, n = 79 variants. h) Spearman’s correlation between validation growth curves in cytosine media and DMS cytosine score, n = 79 variants. i) Spearman’s correlation between validation growth curves in cytosine media and DMS 5-FC + cytosine score, n = 38 variants.