Extended Data Fig. 4: Infection of M. smegmatis TPP mutants by BPs and its derivatives.

A. Growth curves of the different strains incubated with phage BPs∆33HTH_HRM10 or BPs∆33HTH_HRM10-mCherry at MOI 10 or without (UNT) for 2 days in 7H9/OADC supplemented with 1 mM CaCl₂ at 37 °C without agitation. Measurements were taken every 3 hours. Data shown are represented as median of three independent experiments done in triplicate ± interquartile range. Two-sided Dunn’s multiple comparisons test was used to perform statistical analysis. Statistical analysis was done to compare the differences at 48 hours between each strain, p values are mentioned on each plot. B. Representative microscope fields of M. smegmatis strains infected with the fluorophage BPs∆33HTH_HRM10-mCherry (designated BPs mCherry) (MOI 10) for 2 hours at 37 °C. Similar results were obtained at least three times. Scale bars: 30 µm. C. Flow cytometry data plotted as a dot plot showing the percentage of bacilli infected with BPs∆33HTH_HRM10-mCherry relative to the study population. This assay was conducted twice with similar results obtained. D. Phage infection of M. smegmatis TPP mutants. Phages as shown on the left were tenfold serially diluted and spotted onto solid media with M. smegmatis mc²155, M. smegmatis ∆papA3:pLA155 or M. smegmatis ∆papA3:pKSW131. E. Plaquing of BPs∆33HTH_HRM10 and the mCherry derivative on M. smegmatis strains defective in TPP production (left panels) and the complemented strains (right panels). Phage lysates were tenfold serially diluted prior to spotting on bacterial lawns. Similar results were obtained at least three times and a representative experiment is shown.