Fig. 3: Detection and quantification of successful and unsuccessful repair of replication errors.
a Distributions of YFP-MutL foci lifetimes in strains with varying repair efficiency: mutH cells (in blue, 748 foci), mutH cells expressing MutH from the PBAD promoter on a plasmid (mutH pMutH), grown either in LB (mutH pMutH in cyan, 341 foci) or in LB supplemented with 0.05% of arabinose (mutH pMutH ara 0.05% in gray, 1190 foci), and WT cells (in red, 6241 foci). The dashed gray line represents the threshold at 14 min, which separates short-lived foci (on the left) from long-lived foci (on the right). Data presented in the figure are from one representative experiment for each condition. The total number of experiments performed for each condition are: 3 for mutH, WT, and mutH pMutH; 2 for mutH pMutH ara 0.05%. b Estimates of the mutation rate obtained using two approaches: long-lived YFP-MutL foci (lifetime >14 min) (light gray, total number of experiments is the same as in panel (a), and mutation accumulation + whole genome sequencing experiment (MA + WGS, dark gray) for the same conditions as shown in panel (a). The bars represent the average of individual experiments (dots, n = 3 for mutH and mutH pMutH ara 0.05%, n = 4 for mutH pMutH and n = 2 for WT). The values of MA + WGS for the WT strain are from Lee et al.2. c Example of a kymograph of WT cells growing in one microchannel over a 38-min period, showing foci tracks (green and red lines; foci appearing on a single frame are indicated by red arrows). In total 3 WT experiments have been performed with similar results. Source data are provided as a Source Data file.
