Fig. 2.

The monocin element affects ϕ10403S excision within macrophage cells. a Intracellular growth analysis of WT Lm and mutants lacking each of the phage elements, Δmon and Δϕ (monocin cluster and ϕ10403S, respectively), and a double mutant lacking both phage elements Δmon/Δϕ, in BMDM cells. Growth curves represent one biological replicate, more independent experiments are shown in the source file. Error bars represent standard deviation of triplicate samples, and are hidden by the symbols. b A bacterial phagosomal escape assay. Percentage of bacteria that escaped the macrophage phagosomes at 2.5 h post infection, as determined by a microscope fluorescence assay. Macrophages were infected with WT Lm, Δmon and Δmon/Δϕ bacteria, as well as with a Δmon mutant that was complemented with an intact comK gene on the pPL2 plasmid (Δmon+pPL2-comK). The data is a mean of three independent experiments. The error bar represent standard deviation. Asterisk (*) indicates statistical significance of p = 0.01 calculated using Student's t-test. c qRT-PCR analysis of intact comK gene (representing ϕ10403S attB site) in WT Lm and indicated mutants grown intracellularly in BMDM cells (6 h post infection). Presented as relative quantity (RQ), relative to the levels in WT bacteria. The data represent three independent experiments. Error bars indicate a 95% confidence interval. d Intracellular growth analysis of WT Lm, Δmon and the Δmon mutant complemented with an intact comK gene with its native promoter (Δmon+pPL2-comK) in BMDM cells. Growth curves represent one biological replicate, more independent experiments are shown in Supplementary Fig. 2 and in the source data file. Error bars represent standard deviation of triplicate samples, and are hidden by the symbols. Source data are provided as a Source Data file.